Microbiological Control in Poultry Processing

ABSTRACT

In the processing of poultry, equipment, instruments, apparatus and/or water used in such processing, and/or carcasses and/or parts of poultry resulting from the processing of poultry, are disinfected with aqueous solutions of certain halogen-based microbiocides, especially certain bromine-based microbiocides. Described are the particular microbiocides used and the substantial advantages of using such materials, in some cases as concentrated solutions and in other cases as dilute solutions.

REFERENCE TO RELATED APPLICATION

This application is a division of commonly-owned copending applicationSer. No. 11/103,703, filed Apr. 11, 2005, which in turn is acontinuation of commonly-owned application Ser. No. 10/029,329, filedDec. 21, 2001, now U.S. Pat. No. 6,908,636 B2, all disclosure ofwhich—except for the first three paragraphs and the headings thereof inthe specification—is incorporated herein by reference, whichcommonly-owned Application in turn is a continuation-in-part ofcommonly-owned copending application Ser. No. 09/893,581, filed Jun. 28,2001, now abandoned.

REFERENCE TO OTHER COMMONLY-OWNED APPLICATIONS

Reference is hereby made to the following commonly-owned applications:application Ser. No. 09/088,300, filed Jun. 1, 1998, now U.S. Pat. No.6,068,861 issued May 30, 2000; application Ser. No. 09/296,499, filedApr. 22, 1999, now U.S. Pat. No. 6,110,387 issued Aug. 29, 2000;application Ser. No. 09/323,348, filed Jun. 1, 1999, now U.S. Pat. No.6,303,038 B1 issued Oct. 16, 2001; application Ser. No. 09/404,184,filed Sep. 24, 1999, now U.S. Pat. No. 6,322,822 issued Nov. 27, 2001;application Ser. No. 09/442,025, filed Nov. 17, 1999, now U.S. Pat. No.6,306,441 issued Oct. 23, 2001; application Ser. No. 09/451,319, filedNov. 30, 1999; application Ser. No. 09/451,344, filed Nov. 30, 1999, nowU.S. Pat. No. 6,352,725 issued Mar. 5, 2002; application Ser. No.09/456,781, filed Dec. 8, 1999, now U.S. Pat. No. 6,495,169 issued Dec.17, 2002; application Ser. No. 09/483,896, filed Jan. 18, 2000, now U.S.Pat. No. 6,448,410 issued Sep. 10, 2002; application Ser. No.09/484,687, filed Jan. 18, 2000, now U.S. Pat. No. 6,508,954 issued Jan.21, 2003; application Ser. No. 09/484,844, filed Jan. 18, 2000, now U.S.Pat. No. 6,809,205 issued Oct. 26, 2004; application Ser. No.09/484,891, filed Jan. 18, 2000, now U.S. Pat. No. 6,495,698 issued Dec.17, 2002; application Ser. No. 09/484,938, filed Jan. 18, 2000, now U.S.Pat. No. 6,565,868 issued May 20, 2003; application Ser. No. 09/487,816,filed Jan. 18, 2000, now U.S. Pat. No. 6,680,070 issued Jan. 20, 2004;application Ser. No. 09/506,911, filed Feb. 18, 2000, now U.S. Pat. No.6,511,682 issued Jan. 28, 2003; application Ser. No. 09/658,839, filedSep. 8, 2000, now U.S. Pat. No. 6,375,991 issued Apr. 23, 2002;application Ser. No. 09/663,788, filed Sep. 18, 2000, now U.S. Pat. No.6,348,219 issued Feb. 19, 2002; application Ser. No. 09/663,948, filedSep. 18, 2000, now U.S. Pat. No. 6,299,909 B1 issued Oct. 9, 2001;application Ser. No. 09/732,601, filed Dec. 7, 2000, now U.S. Pat. No.6,506,418 issued Jan. 14, 2003; application Ser. No. 09/775,516, filedFeb. 2, 2001, now U.S. Pat. No. 6,641,828 issued Nov. 4, 2003;application Ser. No. 09/778,228, filed Feb. 6, 2001, now abandoned;application Ser. No. 09/785,890, filed Feb. 16, 2001; application Ser.No. 09/893,581, filed Jun. 28, 2001, now abandoned; and application Ser.No. 09/974,622, filed Oct. 9, 2001, now U.S. Pat. No. 6,652,889 issuedNov. 25, 2003.

REFERENCE TO A JOINTLY-OWNED APPLICATION

Reference is hereby made to application Ser. No. 10/028,631, filed Dec.21, 2001, now U.S. Pat. No. 6,919,364 B2.

BACKGROUND

Poultry processing is an area in which microbiological control is ofvital importance. By the very nature of the processing involved thereare numerous opportunities for the poultry to be exposed to variouspathogens in the form of mobile bacteria such as for example Escherichiacoli, Salmonella enteritidis, Salmonella typhimurim, Campylobacterjejuni, Campylobacter coli, Campylobacter lari, and in the form ofbiofilms such as for example Listeria monocytogenes,Pseudomonasfluorescens, Pseudomonas aeruginosa, Enterococcus faecium,and Staphylococcus aureus. The thought of handling, processing andconsuming bacteria-infested poultry is revolting in the extreme.

Heretofore certain chlorine-based microbiocides have been proposed andused in an attempt to provide suitable sanitation in connection withpoultry processing. Unfortunately while some chlorine-basedmicrobiocides show some effectiveness, they possess a number of seriousshortcomings. For one thing they are not as effective as one might wish.Secondly, they tend to be odorous and in many cases can exert ableaching effect upon the poultry carcasses which can prove unpalatableto the consumer. Moreover, because of the spread of fecal matterassociated with the evisceration of the fowl, fecal bacteria abound.This egregious condition in turn results in high nitrogen levels in thewash waters, and on wet surfaces such as cutting surfaces, conduits,tank surfaces, and other downstream equipment exposed one way or anotherto these wash waters. Unfortunately, the active chlorine species ofcertain chlorine-based microbiocides tend to react with the nitrogenousspecies to form chloroamines which are lachrymators as well as beingcorrosive to metallic surfaces. In fact, as little as 50 ppm of chlorinein aqueous washing tanks containing nitrogenous impurities can producequantities of air-borne lachrymators that are intolerable to plantworkers. Furthermore, the consumption of chlorine values in formingchloramines results in a significant loss of biocidal effectivenessinasmuch as the chloroamines are not biocidally-active species.

Clearly therefore a need exists for a new, more effective, economicallyfeasible way of providing microbiological control in the poultryprocessing industry.

BRIEF SUMMARY OF THE INVENTION

This invention fulfills the foregoing need by providing and utilizing incertain highly effective halogen-based microbiocides in the processingof poultry and in the disinfection of equipment, instruments, apparatus,and/or water used in the processing of poultry, and/or of carcassesand/or parts of poultry resulting from the processing of poultry.Microbiocidal agents used pursuant to this invention can be producedeconomically in straightforward processing from relatively low cost rawmaterials and because of their effectiveness, can providemicrobiological control on an economical basis consistent with the needsof the industry.

In one of its embodiments this invention provides in the processing ofpoultry, the improvement which comprises disinfecting equipment,instruments, apparatus and/or water used in such processing, and/orcarcasses and/or other parts of poultry resulting from such processing,with a halogen-based microbiocide which is:

-   (I) an aqueous microbiocidal solution of one or more active halogen    species, which solution is a derivative product in an aqueous medium    of (a) bromine, chlorine, or bromine chloride, or any two or all    three thereof, and (b) a water-soluble source of sulfamate anion; or-   (II) an aqueous microbiocidal solution of one or more active halogen    species, which solution is a derivative product in an aqueous medium    of at least one 1,3-dihalo-5,5-dialkylhydantoin in which one of the    halogen atoms is a chlorine atom and the other is a chlorine or    bromine atom, and in which each of the alkyl groups, independently,    contains in the range of 1 to about 4 carbon atoms; or-   (III) an aqueous microbiocidal solution of one or more active    halogen species, which solution is a derivative product in an    aqueous medium of at least one 1,3-dibromo-5,5-dialkylhydantoin in    which one of the alkyl groups is a methyl group and the other alkyl    group contains in the range of 1 to about 4 carbon atoms: or-   (IV) any two or more of (I), (II), and (III).    The derivative product of (I) above is an aqueous microbiocidal    solution of one or more active halogen species, which solution is    formed by and thus results from a reaction in water between bromine,    chlorine, or bromine chloride, or any two or all three thereof, and    a water-soluble source of sulfamate anion. A concentrated solution    of this type containing over 100,000 ppm of active halogen is    available commercially from Albemarle Corporation under the    trademark STABROM® 909 biocide. A concentrated solution such as this    can be applied to equipment, instruments, or apparatus used in    poultry processing and added to water used in poultry processing    with or without first being further diluted with water. On the other    hand, such a concentrated solution should be diluted with or in    water before application to poultry carcasses or parts thereof, such    as by addition of the concentrate to water in a chilling tank or the    like. Similarly, the derivative products of (II) and (III) above are    aqueous microbiocidal solutions of one or more active halogen    species, which solutions are formed by and thus result from    dissolving the specified 1,3-dihalo-5,5-dialkylhydantoin(s) in    water. Such 1,3-dihalo-5,5-dialkylhydantoins are typically available    commercially in the form of solids and concentrated aqueous    solutions can be formed from such solids for application with or    without further dilution to equipment, instruments, or apparatus    used in poultry processing and added to water used in poultry    processing. But for application to poultry carcasses or parts    thereof, either the concentrated solution should be further diluted    with water before use, or the selected    1,3-dihalo-5,5-dialkylhydantoin solids should be added to water in    proportions yielding the desired microbiocidal dosage directly    without forming an intermediate more concentrated solution.

Purely for convenience, the microbiocides of (I) described above whenmade from bromine chloride, bromine and chlorine, or bromine, chlorine,and bromine chloride, and a sulfamate source, are sometimes referred tohereinafter as “sulfamate-stabilized bromine chloride” even thoughtechnically the actual chemical species in the aqueous medium are mostprobably not bromine chloride molecules or sulfamate adducts orcomplexes of bromine chloride. Thus the designation“sulfamate-stabilized bromine chloride” is simply a shorthand way ofreferring to such compositions, and the designation does not signify,suggest, or imply anything about the actual chemical structure of thecomposition.

In preferred embodiments, the halogen-based microbiocide used in theabove process is (A) a bromine-based microbiocide comprising anoverbased aqueous microbiocidal solution of one or more active brominespecies, said species resulting from a reaction in water between bromineor bromine chloride, a mixture of bromine chloride and bromine, or acombination of bromine and chlorine in which the molar amount ofchlorine is either equivalent to the molar amount of bromine or lessthan the molar amount of bromine, and a water-soluble source ofsulfamate anion, or (B) an aqueous microbiocidal solution of at leastone 1,3-dibromo-5,5-dialkylhydantoin in which one of the alkyl groups isa methyl group and the other alkyl group contains in the range of 1 toabout 4 carbon atoms, or (C) both of (A) and (B) hereof. Thus in theembodiments of this invention wherein equipment, instruments, apparatusand/or water used in poultry processing is disinfected, and/or carcassesand/or other parts of poultry resulting from such processing aredisinfected, “bromine-based” means any of the microbiocides referred toin this paragraph as (A), (B), or (C). In practice, the surfaces to bedisinfected are contacted with the aqueous microbiocidal solutions of(A), (B), or (C) which of course contain a microbiocidally-effectiveamount of the microbiocidal agent and/or microbiocidal hydrolysisproduct(s) thereof.

Such bromine-based microbiocides are more effective than chlorine-basedmicrobiocides against various bacteria and biofilms. In addition, thesebromine-based microbiocides tend to be less odorous than chlorine-basedmicrobiocides, and are essentially devoid of unwanted bleachingactivity. Moreover, while some of the bromine-based microbiocides maypossibly react with nitrogenous species, such as are present in waterand on surfaces associated with poultry processing, the resultantbromamines would also possess microbiological activity. Thus such sidereactions would not materially decrease the microbiologicaleffectiveness made available to the poultry processor by use of thesebromine-based microbiocides. Furthermore, bromamines generally do notexhibit obnoxious properties toward workers in the processing plantwhereas chloramines resulting from use of certain chlorine-basedmicrobiocides under the same conditions tend to be powerfullachrymators.

As noted above, the halogen-based microbiocides of (I) above aremicrobiocidal solutions of one or more active halogen species, whichsolutions are derivative products in a aqueous medium such as water ofbromine, chlorine, or bromine chloride, or any two or all three thereof,and a water-soluble source of sulfamate anion. Likewise, the preferredbromine-based microbiocides of (A) above are microbiocidal solutions ofone or more active bromine species, which solutions are derivativeproducts in a aqueous medium such as water of bromine or brominechloride, a mixture of bromine chloride and bromine, or a combination ofbromine and chlorine in which the molar amount of chlorine is eitherequivalent to the molar amount of bromine or less than the molar amountof bromine, and a water-soluble source of sulfamate anion. To form thesederivative products the components from which the derivative productsare formed are brought together in an aqueous medium such as water,which medium or water, when forming the product, preferably is always ata pH of at least 7 and more preferably is always at a pH higher than 7,e.g., in the range of 10-14, by use of an inorganic base such as sodiumhydroxide. When using a commercially-available product of this type(Stabrom® 909 biocide; Albemarle Corporation), the pH of the aqueousproduct as received is normally in the range of 13 to 14.

Similarly, the halogen-based microbiocides of (II) above aremicrobiocidal solutions of one or more active halogen species, whichsolutions are derivative products in an aqueous medium such as water ofat least one 1,3-dihalo-5,5-dialkylhydantoin in which one of the halogenatoms is a chlorine atom and the other is a chlorine or bromine atom andthe alkyls are as described. Of the halogen-based microbiocides of (II)above, preferred are microbiocidal solutions of one or more activehalogen species, which solutions are derivative products in an aqueousmedium such as water of at least one 1,3-dihalo-5,5-dialkylhydantoin inwhich one of the halogen atoms is a bromine atom and the other is achlorine atom (and the alkyls are as described). The bromine-basedmicrobiocides of (III) above and of (B) above are microbiocidalsolutions of one or more active bromine species, which solutions arederivative products in an aqueous medium such as water of at least one1,3-dibromo-5,5-dialkylhydantoin in which the alkyls are as described.Upon dissolving in an aqueous medium such as water a1,3-dihalo-5,5-dialkylhydantoin referred to in this paragraph, atransformation takes place so that active halogen (or bromine) speciesare present in the resultant solution.

The aqueous microbiocidal solutions used pursuant to the aboveembodiments of this invention can be formed in many cases by adding themicrobiocidal agent itself (i.e., in undiluted form) or as a preformedconcentrated aqueous solution thereof to water being used in one or morepoultry processing operations (e.g., water flowing into chill tanks, orwater already in chill tanks, etc.) to form a diluted microbiocidalsolution of this invention which contacts the surfaces to bedisinfected. Alternatively, a concentrated preformed aqueous solution ofthe microbiocidal agent can be applied directly to the surfaces to bedisinfected (e.g., surfaces of cutting tables, or knives, or etc.), ormore usually such concentrated solution would be mixed with water toform a more dilute solution of the microbiocidal agent which is appliedto the surfaces to be disinfected and/or introduced into water beingused in poultry processing operations. In short, the aqueousmicrobiocidal solutions used pursuant to these embodiments of theinvention can be made in whole or in part from water already in use orto be used in the poultry processing operations, or can be made entirelyfrom water separate from that used or to be used in the poultryprocessing. In each such case, the contacting of the aqueousmicrobiocidal solution however produced and/or applied to the surfacesresults in effective disinfection.

At present the most preferred bromine-based microbiocide used in thepractice of any embodiment of this invention is a water-soluble1,3-dibromo-5,5-dialkylhydantoin in which one of the alkyl groups is amethyl group and the other is an alkyl group containing from 1 to about4 carbon atoms, with 1,3-dibromo-5,5-dimethylhydantoin being the mostpreferred of all.

Various embodiments and features of this invention will be still furtherapparent from the ensuing description and appended claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graphical depiction of the effect of chill tankmicrobiocidal treatments on growth of Pseudomonas species on chickenskin.

FIG. 2 is a graphical depiction of the effect of chill tankmicrobiocidal treatments on growth of total aerobic bacteria on chickenskin.

FIG. 3 is a graphical depiction of the effect of chill tankmicrobiocidal treatments on growth of Pseudomonas species on chickenskin.

FIG. 4 is a graphical depiction of the effect of chill tankmicrobiocidal treatments on growth of total aerobic bacteria on chickenskin.

FIGS. 5 and 6 are graphical depictions of the results obtained in testsinvolving use of bromine species derived from sulfamate-stabilizedbromine chloride in eradicating HPC (heterotrophic plate count) bacteriain biofilm and in planktonic form, respectively, at concentrations inwater of 0.5 ppm and 2 ppm as bromine.

FIGS. 7 and 8 are graphical depictions of the results obtained in testsinvolving use of bromine species derived from sulfamate-stabilizedbromine chloride in eradicating HPC (heterotrophic plate count) bacteriain biofilm and in planktonic form, respectively, at concentrations inwater of 4 ppm and 10 ppm as bromine.

FIG. 9 is a graphical depiction of the results obtained in testsinvolving use of bromine species derived from1,3-dibromo-5,5-dimethylhydantoin in eradicating HPC (heterotrophicplate count) bacteria in a biofilm at concentrations in water of 0.5 and5 ppm as bromine.

FIG. 10 is a graphical depiction of the results obtained in testsinvolving use of bromine species derived from1,3-dibromo-5,5-dimethylhydantoin in eradicating planktonic HPC(heterotrophic plate count) bacteria at concentrations in water of 0.5and 5 ppm as bromine.

FURTHER DETAILED DESCRIPTION OF THE INVENTION

One group of halogen-based microbiocides for use in disinfection ofequipment, instruments, apparatus, and/or water used in the processingof poultry, and/or of carcasses and/or parts of poultry resulting fromthe processing of poultry pursuant to this invention is an aqueousmicrobiocidal solution of one or more active halogen species, saidspecies resulting from a reaction in water between bromine, chlorine, orbromine chloride, or any two or all three thereof, and a water-solublesource of sulfamate anion. If sulfamic acid is used in forming thismicrobiocide, the solution should also be provided with a base,preferably enough base to keep the solution alkaline, i.e., with a pHabove 7, preferably above about 10 and most preferably about 13 orabove. The lower the pH, the more unstable the solution, and thus if thesolution is prepared on site for immediate use, the use of a base is notessential. However, it is preferable to employ a concentratedmicrobiocidal solution manufactured elsewhere, and in such case theconcentrated solution would be provided as an overbased solution with apH of, say, about 13 or more. Often such concentrated solutions willcontain over 50,000 ppm (wt/wt) of active halogen, preferably at leastabout 100,000 ppm (wt/wt) of active halogen, and sometimes as much asabout 150,000 ppm (wt/wt) or more of active halogen, active halogencontent being determinable by use of conventional starch-iodinetitration.

One preferred group of this type is a bromine-based microbiocidalsolution formed by reacting bromine or, more preferably brominechloride, a mixture of bromine chloride and bromine, or a combination ofbromine and chlorine in which the molar amount of chlorine is eitherequivalent to the molar amount of bromine or less than the molar amountof bromine, in an aqueous medium with sulfamic acid and/or awater-soluble salt of sulfamic acid. Except when made on site forimmediate use, such solutions should be highly alkaline solutionstypically with a pH of at least about 12 and preferably at least about13, such pH resulting from use of a base such as sodium hydroxide or thelike, in producing the solution. Concentrated solutions of this type areavailable in the marketplace, for example, Stabrom® 909 biocide(Albemarle Corporation). Processes for producing these concentratedaqueous microbiocidal solutions are described in commonly-owned U.S.Pat. Nos. 6,068,861, issued May 30, 2000, and 6,299,909 B1, issued Oct.9, 2001, all disclosures of which are incorporated herein by reference.

It will be appreciated that even where the microbiocide is made frombromine chloride, a mixture of bromine chloride and bromine, or acombination of bromine and chlorine in which the molar amount ofchlorine is either equivalent to the molar amount of bromine or lessthan the molar amount of bromine is used, the microbiocide isbromine-based as most of the chlorine usually winds up as a chloridesalt such as sodium chloride since an alkali metal base such as sodiumhydroxide is typically used in the processing to raise the pH of theproduct solution to at least about 13. Thus the chlorine in the productsolution is not present as a significant microbiocide.

Another group of halogen-based microbiocides for use in theseembodiments of this invention is one or moreN,N′-halo-5,5-dialkylhydantoins in which one of the halogen atoms ischlorine and the other is bromine or chlorine, and in which the alkylgroups, independently, each contain from 1 to about 4 carbon atoms.Suitable compounds of this type include, for example, such compounds as1,3-dichloro-5,5-dimethylhydantoin, 1,3-dichloro-5,5-diethylhydantoin,1,3-dichloro-5,5-di-n-butylhydantoin,1,3-dichloro-5-ethyl-5-methylhydantoin,N,N′-bromochloro-5,5-dimethylhydantoin,N,N′-bromochloro-5-ethyl-5-methylhydantoin,N,N′-bromochloro-5-propyl-5-methylhydantoin,N,N′-bromochloro-5-isopropyl-5-methylhydantoin,N,N′-bromochloro-5-butyl-5-methylhydantoin,N,N′-bromochloro-5-isobutyl-5-methylhydantoin,N,N′-bromochloro-5-sec-butyl-5-methylhydantoin,N,N′-bromochloro-5-tert-butyl-5-methylhydantoin,N,N′-bromochloro-5,5-diethylhydantoin, and mixtures of any two or moreof the foregoing. N,N′-bromochloro-5,5-dimethylhydantoin is availablecommercially under the trade designation Bromicide® biocide (Great LakesChemical Corporation). Another suitable bromochlorohydantoin mixture iscomposed predominantly of N,N′-bromochloro-5,5-dimethylhydantointogether with a minor proportion by weight of1,3-dichloro-5-ethyl-5-methylhydantoin. A mixture of this latter type isavailable in the marketplace under the trade designation Dantobrom®biocide (Lonza Corporation).

When a mixture of two or more of the foregoingN,N′-bromochloro-5,5-dialkylhydantoin biocides is used pursuant to thisinvention, the individual biocides of the mixture can be in anyproportions relative to each other.

It will be understood that the designation N,N′ in reference to, say,N,N′-bromochloro-5,5-dimethylhydantoin means that this compound can be(1) 1-bromo-3-chloro-5,5-dimethylhydantoin, or (2)1-chloro-3-bromo-5,5-dimethylhydantoin, or (3) a mixture of1-bromo-3-chloro-5,5-dimethylhydantoin and1-chloro-3-bromo-5,5-dimethylhydantoin. Also, it is conceivable thatsome 1,3-dichloro-5,5-dimethylhydantoin and1,3-dibromo-5,5-dimethylhydantoin could be present in admixture with(1), (2), or (3).

An even more preferred system for use in the practice of theseembodiments of this invention is a bromine-based microbiocidal solutionof a 1,3-dibromo-5,5-dialkylhydantoin in which one of the alkyl groupsis a methyl group and the other alkyl group contains in the range of 1to about 4 carbon atoms. Thus these preferred biocides comprise1,3-dibromo-5,5-dimethylhydantoin,1,3-dibromo-5-ethyl-5-methylhydantoin,1,3-dibromo-5-n-propyl-5-methylhydantoin,1,3-dibromo-5-isopropyl-5-methylhydantoin,1,3-dibromo-5-n-butyl-5-methylhydantoin,1,3-dibromo-5-isobutyl-5-methylhydantoin,1,3-dibromo-5-sec-butyl-5-methylhydantoin,1,3-dibromo-5-tert-butyl-5-methylhydantoin, and mixtures of any two ormore of them. Of these biocidal agents,1,3-dibromo-5-isobutyl-5-methylhydantoin,1,3-dibromo-5-n-propyl-5-methylhydantoin, and1,3-dibromo-5-ethyl-5-methylhydantoin are, respectively, preferred, morepreferred, and even more preferred members of this group from the costeffectiveness standpoint. Of the mixtures of the foregoing biocides thatcan be used pursuant to this invention, it is preferred to use1,3-dibromo-5,5-dimethylhydantoin as one of the components, with amixture of 1,3-dibromo-5,5-dimethylhydantoin and1,3-dibromo-5-ethyl-5-methylhydantoin being particularly preferred. Themost preferred member of this group of microbiocides is1,3-dibromo-5,5-dimethylhydantoin. This compound is available in themarketplace in tablet or granular form under the trade designationsAlbrom® 100T biocide and Albrom® 100PC biocide (Albemarle Corporation).

When a mixture of two or more of the foregoing1,3-dibromo-5,5-dialkylhydantoin biocides is used pursuant to thisinvention, the individual biocides of the mixture can be in anyproportions relative to each other.

Methods for producing 1,3-dibromo-5,5-dialkylhydantoins are known andreported in the literature.

If desired, the 1,3-dihalo-5,5-dialkylhydantoins can be dissolved in asuitable innocuous, harmless, water-soluble organic solvent with orwithout water to form a solution which can be applied to surfaces ofequipment, instruments, or apparatus. Depending upon the solvent used,the surfaces can then be further washed with clean water to removeresidues from such solvent. Besides increasing the amount of1,3-dihalo-5,5-dialkylhydantoin that can be put into solution thusfacilitating formation of a concentrated solution, e.g., on the premisesof the poultry processing, such a concentrated solution when dilutedsuch as by addition to process water being used on the premisespossesses microbiocidal activity from the1,3-dihalo-5,5-dialkylhydantoin. Thus aqueous solutions used pursuant tothis invention can contain suitably small amounts of an innocuous,harmless, water-soluble organic solvent, which non-toxic, at least atthe dosage levels involved, such as acetonitrile.

In cases where extremely powerful biocidal activity is desired such asduring periodic cleaning and disinfection operations, concentratedaqueous solutions of the microbiocides of this invention can be directlyapplied to surfaces of poultry processing equipment, instruments and/orapparatus infested with pathogenic microorganisms. Such concentratedsolutions can contain, for example, as much as 150,000 ppm or 160,000ppm or more of active bromine, and as much as about 66,667 ppm or about71,111 ppm of active chlorine, as determinable by conventionalstarch-iodine titration. If desired, a portion of such concentratedsolution can be diluted with any suitable amount of water beforeapplication directly to the surfaces of such poultry processingequipment, instruments and/or apparatus, provided of course that thediluted solution still contains a microbiocidally-effective amount ofactive bromine species for the use at hand. Also, concentrated solutionsof this invention can be added to and thus used in diluted form inprocess water being used in poultry processing operations, such as forexample, in water flowing through conduits, in water flowing into orbeing maintained in tanks, and in water being used in sprayingequipment.

The amount (concentration) of the selected microbiocide utilized in thepractice of this invention will vary depending on various factors suchas the particular microbiocide being used, the nature and frequency ofprior microbiocidal treatments, the types and nature of themicroorganisms present, the amount and types of nutrients available tothe microorganisms, the nature and extent of cleansing actions, if any,taken in conjunction with the microbiocidal treatment, the surface orlocus of the microorganisms being treated, and so on. In any event, amicrobiocidally-effective amount of the diluted aqueous solution of themicrobiocide of this invention will be applied to or contacted with themicroorganisms. Typically the diluted solution will contain amicrobiocidally-effective amount of active halogen in the range of about2 to about 1000 ppm (wt/wt), preferably in the range of about 2 to about500 ppm (wt/wt), and more preferably in the range of about 25 to about250 ppm (wt/wt), active halogen being determinable by use of theconventional DPD test procedure. If the actual active halogen in thesolution consists of active chlorine, the concentration of the dilutedsolution used is preferably at least two to three times higher than theminimums of the foregoing ranges. In the case of the1,3-dibromo-5,5-dialkylhydantoins used pursuant to this invention, aparticularly preferred range for use in ordinary situations (e.g.,washing hard surfaces such as tables, walls, floors, conveyor machineryor parts thereof such as converor belts or shackles, and knives orcutting blades) is in the range of about 50 to about 150 ppm (wt/wt) ofactive bromine. When contacting poultry carcasses or edible partsthereof with aqueous solutions formed from at least one1,3-dibromo-5,5-dialkylhydantoin used pursuant to this invention, it isespecially preferred to use in the water for washing or otherwisecontacting the poultry carcasses or edible parts thereof, amicrobiocidally effective amount of active bromine that does notsignificantly or appreciably bleach the skin of the caracass or have asignificant or appreciable adverse effect upon the organleptic taste ofcooked meat from the poultry such as the breast meat and thigh meat.Such amount is typically within the range of about 0.5 to about 30 ppm(wt/wt) and preferably in the range of about 5 to about 25 ppm (wt/wt)of active bromine as determinable by the DPD test procedure. Similarranges are deemed applicable if using sulfamate-stabilized brominechloride in these carcass washing operations. It will be understood thatdepartures from the foregoing ranges can be made whenever deemednecessary or desirable, and such departures are within the spirit andscope of this invention.

Consequently, depending upon the way in which the microbiocide of thisinvention is being used, a microbiocidally-effective amount of themicrobiocides of this invention can extend from as little as about 2 ppmup to as high as the maximum water solubility of the particular activehalogen microbiocidal agent being used, at the temperature at which suchactive halogen microbiocidal agent is being used.

As can be seen from the above, there are two different types ofprocedures that are used for determining active halogen content, whetheractive chlorine, active bromine or both. For measuring concentrations inthe vicinity of above about, say, 500 ppm or so (wt/wt) of activebromine or, say, above about 1100 ppm of active chlorine, starch-iodinetitration is the preferred procedure. On the other hand, whereconcentrations are below levels in these vicinities, the conventionalDPD test procedure is more suitable, as this test is designed formeasuring very low active halogen concentrations, e.g., active chlorineconcentrations in the range of from zero to about 11-12 ppm (wt/wt) oractive bromine concentrations in the range of from zero to about 5 ppm(wt/wt). In fact, where the actual concentration of active chlorine isbetween, say, about 11-12 ppm and about 1100 ppm (wt/wt), or the wherethe actual concentration of active bromine is between, say, about 5 ppmand about 1100 ppm (wt/wt), the test sample is typically diluted withpure water to reduce the actual concentration to be in the range ofabout 4 to about 11-12 ppm in the case of active chlorine and to be inthe range of about 2 to about 5 ppm in the case of active bromine beforemaking the DPD analysis. It can be seen therefore that while there is nocritical hard-and-fast concentration dividing line between whichprocedure to use, the approximate values given above represent apractical approximate dividing line, since the amounts of water dilutionof more concentrated solutions when using the DPD test procedureincrease with increasing initial active halogen concentration, and suchlarge dilutions can readily be avoided by use of starch-iodine titrationwhen analyzing the more concentrated solutions. In short, with suitablydilute solutions use of the DPD test procedure is recommended, and withmore concentrated solutions use of starch-iodine titration isrecommended.

The starch-iodine titration procedure for determination of activehalogen has long been known. For example, chapter XIV of Willard-Furman,Elementary Quantitative Analysis, Third Edition, D. Van NostrandCompany, Inc., New York, Copyright 1933, 1935, 1940 provides adescription of starch-iodine titration. While details of standardquantitative analytical procedures for determination of active halogenin such product solutions by starch-iodine titration may vary from caseto case, the results are normally sufficiently uniform from one standardprocedure to another as not to raise any question of unreliability ofthe results. A recommended starch-iodine titration procedure is asfollows: A magnetic stirrer and 50 milliliters of glacial acetic acidare placed in an iodine flask. The sample (usually about 0.2-0.5 g) forwhich the active halogen is to be determined is weighed and added to theflask containing the acetic acid. Water (50 milliliters) and aqueouspotassium iodide (15%, wt/wt; 25 milliliters) are then added to theflask. The flask is stoppered using a water seal. The solution is thenstirred for fifteen minutes, after which the flask is unstoppered andthe stopper and seal area are rinsed into the flask with water. Anautomatic buret (Metrohm Limited) is filled with 0.1 normal sodiumthiosulfate. The solution in the iodine flask is titrated with the 0.1normal sodium thiosulfate; when a faint yellow color is observed, onemilliliter of a 1 wt % starch solution in water is added, changing thecolor of the solution in the flask from faint yellow to blue. Titrationwith sodium thiosulfate continues until the blue color disappears. Theamount of active halogen is calculated using the weight of the sampleand the volume of sodium thiosulfate solution titrated. In this way, theamount of active halogen such as active chlorine or active bromine in anaqueous product solution, regardless of actual chemical form, can bequantitatively determined.

The standard DPD test for determination of low levels of active halogenis based on classical test procedures devised by Palin in 1974. See A.T. Palin, “Analytical Control of Water Disinfection With SpecialReference to Differential DPD Methods For Chlorine, Chlorine Dioxide,Bromine, Iodine and Ozone”, J. Inst. Water Eng., 1974, 28, 139. Whilethere are various modernized versions of the Palin procedures, therecommended version of the test is fully described in Hach WaterAnalysis Handbook, 3rd edition, copyright 1997. The procedure for “totalchlorine” (i.e., active chlorine) is identified in that publication asMethod 8167 appearing on page 379, Briefly, the “total chlorine” testinvolves introducing to the dilute water sample containing activehalogen, a powder comprising DPD indicator powder, (i.e.,N,N′-diethyldiphenylenediamine), KI, and a buffer. The active halogenspecies present react(s) with KI to yield iodine species which turn theDPD indicator to red/pink. The intensity of the coloration depends uponthe concentration of “total chlorine” species (i.e., active chlorine”)present in the sample. This intensity is measured by a calorimetercalibrated to transform the intensity reading into a “total chlorine”value in terms of mg/L Cl₂. If the active halogen present is activebromine, the result in terms of mg/L Cl₂ is divided by 2.25 to expressthe result in terms of mg/L Br₂ of active bromine.

In greater detail, the DPD test procedure is as follows:

-   1. To determine the amount of species present in the water which    respond to the “total chlorine” test, the water sample should be    analyzed within a few minutes of being taken, and preferably    immediately upon being taken.-   2. Hach Method 8167 for testing the amount of species present in the    water sample which respond to the “total chlorine” test involves use    of the Hach Model DR 2010 calorimeter. The stored program number for    chlorine determinations is recalled by keying in “80” on the    keyboard, followed by setting the absorbance wavelength to 530 nm by    rotating the dial on the side of the instrument. Two identical    sample cells are filled to the 10 mL mark with the water under    investigation. One of the cells is arbitrarily chosen to be the    blank. To the second cell, the contents of a DPD Total Chlorine    Powder Pillow are added. This is shaken for 10-20 seconds to mix, as    the development of a pink-red color indicates the presence of    species in the water which respond positively to the DPD “total    chlorine” test reagent. On the keypad, the SHIFT TIMER keys are    depressed to commence a three minute reaction time. After three    minutes the instrument beeps to signal the reaction is complete.    Using the 10 mL cell riser, the blank sample cell is admitted to the    sample compartment of the Hach Model DR 2010, and the shield is    closed to prevent stray light effects. Then the ZERO key is    depressed. After a few seconds, the display registers 0.00 mg/L Cl₂.    Then, the blank sample cell used to zero the instrument is removed    from the cell compartment of the Hach Model DR 2010 and replaced    with the test sample to which the DPD “total chlorine” test reagent    was added. The light shield is then closed as was done for the    blank, and the READ key is depressed. The result, in mg/L Cl₂ is    shown on the display within a few seconds. This is the “total    chlorine” level of the water sample under investigation.

In the practice of this invention the microbiocidal system can be usedin various ways. For example, a microbiocidally effective amount of amicrobiocide of this invention, preferably a bromine-based microbiocidalsystem, is applied to the locus of the microorganisms to be eradicatedor controlled so that the microbiocidal system comes in contact withthese microorganisms. The application can be made by thoroughapplication by pouring, spraying, wet mopping, flooding, and/or wetwiping infested or potentially infested surfaces or areas of theprocessing equipment and environs such as flooring, walls, tables,conveyors, stanchions, conduits, tanks, and drains with abiocidally-effective amount of an aqueous solution the microbiocide.Where applicable and possible, portions of the processing apparatus canbe immersed in an aqueous solution of the microbiocide, with temporarydisassembly, if necessary. Such applications should be conductedroutinely on a frequency sufficient to ensure that exposure of thepoultry being processed to dangerous microorganisms, such as bacteriaand biofilms is prevented to the greatest extent possible. For bestresults these operations should be conducted in conjunction orassociation with thorough cleaning operations such as scrubbing,scouring, scraping and, otherwise removing infestations of biofouling orbiofilms, whether visible or invisible. After contacting themicroorganisms with the microbiocide for a suitable period of time toensure penetration into polysaccharide slimes and other defensemechanisms of various species of these microorganisms, the entiredisinfected area should be washed, e.g., hosed down, with clean waterand preferably the washings themselves should be disinfected withadditional microbiocide of this invention, preferably a bromine-basedmicrobiocide, before discharge. The contact times will of course varydepending upon the frequency and thoroughness of the cleaning anddisinfection operations and the identity and concentration of theparticular microbiocidal solution being employed. Generally speakingcontact times may fall in the range of from about a few minutes to a fewhours, but any period of time that effects the eradication or control ofthe microbial population in the poultry processing areas should be usedand is within the scope of this invention.

Another mode of applying the microbiocidally-effective amounts ofsolid-state microbiocides of these embodiments of the invention is tocause the microbiocide to be leached into water streams passing throughconduits and into tanks or other washing devices utilized in theprocessing of the poultry. For example, suitable solid forms of themicrobiocide, preferably a bromine-based microbiocide, such as tablets,briquettes, pellets, nuggets, or granules are placed in suitable feedingdevices through which a stream of water is passed. The passage of thewater through the bed of the microbiocide results in the streamcontinuously dissolving small quantities of the microbiocide to therebyprovide microbiocidally effective amounts of the microbiocide in thewater. 1,3-Dibromo-5,5-dimethylhydantoin is especially preferred for usein this mode of application because of its relatively low solubility andthus relatively slow rate of dissolution in water at ambient roomtemperatures. This translates into relatively long periods of use beforeneed of refilling the device holding the solids. By way of example, thesolubility of 1,3-dibromo-5,5-dimethylhydantoin in water at 75° F. (ca.24° C.) is 405 ppm expressed as Cl₂ whereas the solubilities ofN,N′-bromochloro-5,5-dimethylhydantoin and of the commercial mixture ofN,N′-bromochloro-5,5-dimethylhydantoin and1,3-dichloro-5-ethyl-5-methylhydantoin at the same temperature are,respectively, 890 ppm and 1905 ppm, both expressed as Cl₂.

An especially cost-effective, operationally efficient, and highlypreferred way of forming aqueous microbiocidal solutions of one or more1,3-dibromo-5,5-dialkylhydantoins in which one of the alkyl groups is amethyl group and the other alkyl group contains in the range of 1 toabout 4 carbon atoms, most preferably 1,3-dibromo-5,5-dimethylhydantoin,(“dibromodialkylhydantoin(s)”) comprises passing water through a bed ofone or more such dibromodialkylhydantoin(s) in granular, nugget, pellet,tablet or other non-powdery particulate form (“bed”) disposed in acanister, tank, or other similar vessel (“tank”). Preferably the tankhas a pressure sealable port at its upper portion for periodicallyreplenishing the contents of the bed, and the water is caused to flowupwardly through a portion of the bed. More preferably, the tank iselongated in an upward direction so that the bed is longer from top tobottom than from side to side, this upward water flow is dispensed intothe bed to flow upwardly through only a lower portion of the bed, andthence substantially horizontally through a port disposed between thelower and the upper portions of the bed and tank. In this way the upperportion of the bed serves as a reserve supply of contents of the bedwhich automatically feeds into the lower portion of the bed undergravity as the lower portion of the bed is slowly but substantiallyuniformly dissolved away in the water flow. Thus in this operation thewater flow is preferably at least a substantially continuous flow, andmost preferably, is a continuous flow. Methods for producing granules,tablets or other non-powdery particulate forms of1,3-dibromo-5,5-dimethylhydantoin are described in detail incommonly-owned copending applications PCT/US01/01541, 01/01545, and01/01585, all filed Jan. 17, 2001, each claiming priority based onrespective earlier-filed corresponding U.S. applications. Excellentprocess technology for producing 1,3-dibromo-5,5-dimethylhydantoin foruse in making such granules, tablets or other non-powdery particulateforms is described in detail in commonly-owned copending applicationPCT/US01/01544, filed Jan. 17, 2001, claiming priority based on anearlier-filed corresponding U.S. application. The disclosures of eachsuch PCT and U.S. application is incorporated herein by reference.Particularly preferred apparatus for use in conjunction with suchgranules, tablets or other non-powdery particulate forms of thesedibromodialkylhydantoin(s) in forming aqueous microbiocidal solutionsthereof is available from Neptune Chemical Pump Company, a division ofR.A. Industries, Inc., Lansdale, Pa. 19446, as “Bromine Feeders” ModelsBT-15, BT-40, BT-42, BT-80, BT-160, BT-270, and BT-350, or equivalent.Excellent results are achieved using combinations of Model BT-40 withgranules of 1,3-dibromo-5,5-dimethylhydantoin Albrom® 100 biocideavailable from Albemarle Corporation. Single charges of suchmicrobiocides in tablet or granular form in such device can providecontinuous highly-effective microbiocidal activity in bodies of end usewater at ordinary outdoor temperatures for as long as five (5) monthswithout need for replenishment.

In the case of the more water-soluble microbiocides used pursuant tothis invention, another suitable method of effecting contact between themicrobiocide and the microorganisms is to pump an aqueous solutioncontaining a microbiocidally-effective amount of the microbiocidethrough the conduits and into the tanks or other washing devices, suchas scalding tanks and chill tanks, utilized in the processing of thepoultry. Variants of this procedure include dispensing portion-wise asby gravity dripping an aqueous solution of the microbiocide directlyinto a tank or other vessel in which poultry are to be or are beingprocessed.

A further mode of application pursuant to these embodiments of theinvention involves applying to or contacting the poultry itself,typically promptly before and/or after slaughter, with an aqueoussolution of the microbiocide. After providing a suitable contact time toeradicate bacteria on the surfaces of the poultry, the poultry can thenbe washed down to remove both the excess microbiocide and the dispatchedmicrobial population from the exposed surfaces of the fowl itself. Theinternal organs of the fowl after slaughter can also be treated andwashed down in the same manner. The application(s) of the microbiocidalsolution(s) in this manner can take any suitable form, e.g., use ofaqueous sprays containing a microbiocidally-effective amount of themicrobiocide being used, or immersion of the fowl or internal organsthereof in one or more tanks containing aqueous solutions ofmicrobiocidally-effective amounts of the microbiocide being used.

Preferably two or more of the foregoing methods of application of themicrobiocides of this invention are used. Thus in a preferred embodimenta microbiocide of these embodiments of the invention, preferably anaqueous bromine-based microbiocidal solution, is applied by (i)periodically contacting at least portions, if not all, of the poultryprocessing apparatus to disinfection or sanitization with amicrobiocidally-effective amount of an aqueous solution of at least onemicrobiocide of these embodiments of the invention, preferably abromine-based microbiocide, and (ii) contacting the exposed surfaces ofthe poultry with a microbiocidally-effective amount of an aqueoussolution of at least one microbiocide of these embodiments of theinvention, preferably a solution of a bromine-based microbiocide, beforeand/or after, preferably after, dispatching the fowl. In anotherpreferred embodiment, a microbiocide of these embodiments of theinvention, preferably an aqueous bromine-based microbiocidal solution,is applied by (i) periodically contacting at least portions, if not all,of the poultry processing apparatus to disinfection or sanitization witha microbiocidally-effective amount of an aqueous solution of at leastone microbiocide of these embodiments of the invention, preferably abromine-based microbiocide, and (ii) contacting the edible portionsand/or internal organs of the dispatched fowl with amicrobiocidally-effective amount of an aqueous solution of at least onemicrobiocide of these embodiments of the invention, preferably asolution of a bromine-based microbiocide.

Particularly preferred processes of this invention are those wherein thefowl are processed by a series of steps which comprise the following:(a) suspending the fowl in moving clamps or shackles, (b) stunning, butnot killing, the fowl such as by use of a suitable gas, or by contactingat least the heads of the fowl with a water-applied electric shock tostun the fowl, e.g., by immersing the heads in a water bath carrying asuitable current to effect the stunning, (c) cutting the jugular veinsand/or carotid arteries at the neck of the stunned fowl either manuallywith a knife or automatically with a mechanical cutting device, (d)draining blood from the carcasses, (e) scalding the birds with hotwater, e.g., in a scalding tank, to facilitate feather removal, (f)defeathering the fowl, (g) removing the heads and feet from the fowl,(h) eviscerating the fowl either manually with a knife, or automaticallywith mechanical evisceration apparatus, (i) separating the viscera fromthe carcasses, (j) washing the carcasses, and (k) chilling thecarcasses, e.g., in water such as by passage of the carcasses through atleast one and often two chill tanks, or by air chilling. The scaldingstep will typically be conducted at water temperatures in the range ofabout 50 to about 60° C., with the lower temperatures being preferredfor retention of normal yellow-colored skin. The higher temperatureswill more usually be used in connection with turkeys and spent egg-layerhens. The chilling temperatures used will typically reduce the carcasstemperature to below about 4° C., with final temperatures of thefinished carcasses for shipment being as low as about −2° C. Other stepscan be included and in some cases one or more of the steps (a) through(j) may be altered or revised or the sequence of the steps may to someextent be altered or revised, to adapt to given circumstances. Examplesof extra steps that may be included are inspection steps, e.g., bygovernmental regulatory personnel, and wax-dipping in the case of waterfowl to enhance the extent of defeathering. Inspections are oftenconducted subsequent to the evisceration step, such as before separatingthe viscera from the carcasses. Wax dipping will typically be used whenprocessing waterfowl, the feathers of which typically are more difficultto remove than, say, chickens. Wax dipping will typically be performeddirectly after use of feather-picking machines which utilize rubber“fingers” to beat off the feathers. The wax dipping step will typicallyinvolve dipping the partially defeathered carcass into a molten waxcontained in a tank, allowing the wax to harden on the carcass, and thenremoving the wax coating as by peeling it off along with feathersembedded in the wax. This operation can be repeated as desired, beforeproceeding to the next step in the process, e.g., removal of the headsand feet. One illustrative example of a suitable revision of thesequence of steps, would be to conduct step (g) before step (d) insteadof after step (f). Upon a reading of this disclosure, other suitablesequence revisions may well become obvious to one of ordinary skill inthe art, and thus need not be further elaborated upon here.

In the above processing, the microbiocidal action of the microbiocidesof these embodiments of the invention, preferably one or more applicablebromine-based microbiocides used pursuant to this invention, can beapplied at any of a variety of suitable stages in the operation. Forexample. an applicable microbiocidal solution of this invention can beapplied to any or all of the processing equipment used including knives,conveying apparatus, the surfaces of emptied scaling tanks, defeatheringapparatus (e.g., rubber “fingers” etc.), knives and mechanical apparatusused for cutting or eviscerating the fowl, all surfaces that come incontact with the blood or the viscera of the fowl, including tables,conveyor belts, etc., and all surfaces that come in contact with thecarcasses after separation of the viscera therefrom. The applicablesanitizing solutions of this invention can be applied to by immersion,spraying, flooding, or any other way of ensuring that themicrobiocidally-effective solution contacts the surfaces that contain orare exposed to the undesirable microorganisms such as bacteria and/orbiofilm (biofouling).

Another way by which, in the above processing, the microbiocidal actionof the applicable microbiocides of this invention, preferably one ormore applicable bromine-based microbiocides used pursuant to thisinvention, can be applied involves including a microbiocidally-effectiveamount of the microbiocide to the water being used at one or more stagesof the processing. Thus the water in the scalding tank(s) and/or in thechill tank(s) can be so treated. Another mode is to include amicrobiocidally-effective amount of the microbiocide to the water usedin washing the carcasses and the viscera at various points where theseparts are handled, separated, and/or processed. The dosage levels atthese different points in the processing can be the same or different asdeemed necessary or desirable.

The practice and advantages of this invention are illustrated by thefollowing non-limiting Examples.

EXAMPLE 1

Comparative tests were conducted to determine the effect on poultrycarcass bacteria (Escherichia coli field strain) during a normal1,5-hour chill tank immersion in water containing differentmicrobiocidal compositions. The effect of these treatments on theresidual chill tank water was also investigated. Carcasses were firstimmersed in a warm bath containing 10⁴ E coli per mL of liquid.Carcasses were then immersed in chill tanks containing normal organicfluids (blood, fat, skin, and meat particles) and containing one of therespective microbiocidal compositions under test. Total bacteria countof whole bird (both inside and outside) was used to determine efficacyof various microbiocidal compositions. The microbiocidal compositionstested were Aquatize® biocide (Bioxy Incorporated, 3733 National Drive,Suite 115, Raleigh, N.C. 27612-4845), sodium hypochlorite (Clorox®bleach), sodium bromide (supplied as a 40% solution in water),combinations of sodium hypochlorite and sodium bromide, and aconcentrated alkaline aqueous solution produced from bromine chlorideand sulfamate anion (SSBC) (Stabrom® 909 biocide; AlbemarleCorporation).

The trial events and experimental design used were as follows:

-   a) For all treatments a total of 190 birds were processed in a    normal fashion. Each treatment involved use of 10 birds.-   b) A warm (100° F.) bath was prepared containing 5×10⁴ DelMarVa    (Delaware-Maryland farm area) field Escherichia coli stain bacteria    per mL. At least 200 mL of total bath fluid was provided for each    bird.-   c) All birds (both controls and treated) were randomly immersed into    the warm bath. Both the inside and outside carcass areas were    immersed to assure complete coverage.-   d) Each separate chill tank water solution (normal tap water    adjusted pH to 8.5, ice added to produce temperatures of <45° F.)    contained 2×10³ per mL bacteria.-   e) The chill tank water solutions (at least 750 mL each) were be    prepared for each disinfecting treatment, and birds were immersed    for a period of 1.5 hours.-   f) Each 10 minutes during the 1.5 hour chilling period, the birds    were completely lifted out of solution and then reimersed in the    solution. After the 1.5 hour chilling period, the birds were taken    from the chill water and drained for 30 seconds, then promptly    (within 5 minutes) placed into a sterile whole bird stomacher bag    containing 400 mL diluent (Butterfield's Phosphate Diluent) bacteria    collection.-   g) Diluent (400 mL) was added to each carcass contained in a sterile    stomacher bag while making sure to pour the diluent into the inside    of the abdominal cavity. The carcass was rinsed inside and out with    a rocking motion for one minute (ca. 35 RPM). This was best done by    grasping the broiler carcass with one hand and the closed top of the    bag with the other then rocking with a reciprocal motion in a 18-24    inch arc, assuring that all carcass surfaces (interior and exterior)    were rinsed.-   h) The rinse solutions were then transferred from each stomacher bag    into individual sample bottles, taking care to ensure that the    information on the date of collection, time of collection, and    treatment group matched that of the sample. Each bottle was sealed    with parafilm and stored in a refrigerator.-   i) Dilution of the fluids was conducted such that a 25-250 count was    present on the MacConkey plate. After the fluids had been diluted,    0.1 mL of fluid was be placed on a MacConkey agar plate and bacteria    counts determined. In cases where the goal of a 25-250 count on the    plate was not achieved, another dilution and replating were    conducted.-   j) After all carcasses for each treatment had been dipped, water    sample bacteria were determined.-   k) Total bacteria per bird were calculated.    The chill water used was composed per liter of 950 mL of tap water,    50 mL of blood, 10 g of ground abdominal fat, and 10 g of meat    particles. To form the bacteria culture used as the test bacteria    source, an overnight culture in BHI broth was transferred to fresh    BHI broth and incubated at 37° C. for 1.5 hour to a population    density of approximately 8×10⁶ DFU per mL (optical density at 600    nm, ˜0.1). This bacteria solution was added equally at 5×10⁴ to    provide a water solution to predip all birds prior to chilling. In    addition, prior to dipping the birds for the 1,5-hour chill period,    additional bacteria were added to the chill water in the amount of    2×10³ total bacteria per mL of chill water. Poultry carcass    microbial contamination measurement was achieved by the complete    washing of the entire carcass surface (inside and outside) using    suitable sterile stripping solution followed by collection and    plating of the stripping solution for bacterial enumeration.

Table 1 presents the experimental design of this group of tests. TABLE 1Test Group Test Material & Disinfectant Level 1 No disinfectant¹ 2Aquatize ® biocide (dilution 1:700), contains 50 ppm sodium chlorite 3Aquatize ® biocide (dilution 1:350), contains 100 ppm sodium chlorite 4Aquatize ® biocide (dilution 1:250), contains 150 ppm sodium chlorite 5Clorox ® bleach 12.5% Cl (dilution 1:2,500), Contains 50 ppm Cl₂equivalent² 6 Clorox ® bleach 12.5% Cl (dilution 1:1,250), Contains 100ppm Cl₂ equivalent 7 Clorox ® bleach 12.5% Cl (dilution 1:800), Contains150 ppm Cl₂ equivalent 8 SSBC (dilution 1:12,500), Contains 50 ppm Cl₂equivalent (1.57 times Cl₂) 9 SSBC (dilution 1:6,250), Contains 100 ppmCl₂ equivalent (1.57 times Cl₂) 10 SSBC (dilution 1:4,000), Contains 150ppm Cl₂ equivalent (1.57 times Cl₂) 11 Bleach and Sodium Bromide (1:1mole ratio mix), bleach dilution 1:3,500 & 40% NaBr solution dilution1:28,000, Contains 50 ppm Cl₂ equivalent (1:1 Cl₂ equivalent) 12 Bleachand Sodium Bromide (1:1 mole ratio mix), bleach dilution 1:1,750 & 40%NaBr solution dilution 1:14,000, Contains 100 ppm Cl₂ equivalent (1:1Cl₂ equivalent) 13 Bleach and Sodium Bromide (1:1 mole ratio mix),bleach dilution 1:1,200 & 40% NaBr solution dilution 1:9,300, Contains150 ppm Cl₂ equivalent (1:1 Cl₂ equivalent) 14 Bleach and Sodium Bromide(2:1 mole ratio mix), bleach dilution 1:3,000 & 40% NaBr solutiondilution 1:50,000, Contains 50 ppm Cl₂ equivalent (1:1 Cl₂ equivalent)15 Bleach and Sodium Bromide (2:1 mole ratio mix), bleach dilution1:1,500 & 40% NaBr solution dilution 1:25,000, Contains 100 ppm Cl₂equivalent (1:1 Cl₂ equivalent) 16 Bleach and Sodium Bromide (2:1 moleratio mix), bleach dilution 1:1,000 & 40% NaBr solution dilution1:16,600, Contains 150 ppm Cl₂ equivalent (1:1 Cl₂ equivalent) 17 SodiumBromide (40% solution), Dilution 1:8,000, Contains 50 ppm Cl₂ equivalent(1:1 Cl₂ equivalent) 18 Sodium Bromide (40% solution), Dilution 1:4,000,Contains 100 ppm Cl₂ equivalent (1:1 Cl₂ equivalent) 19 Sodium Bromide(40% solution), Dilution 1:2,670, Contains 150 ppm Cl₂ equivalent (1:1Cl₂ equivalent)¹Negative Control contained contaminated (bacteria 2.67 × 10⁵ per mL)water.²Positive Control is normal poultry industry practice of adding 50 ppmCl₂ equivalent.

Tables 2-4 show, respectively, the method of determining the dilutionlevels for achieving 50 ppm, 100 ppm, and 150 ppm Cl₂ equivalents in thecase of the chill tank solutions formed from Clorox® bleach solution anda 40% water solution of sodium bromide. TABLE 2 Dilutions for 50 ppm Cl₂Equivalent Percentage (% of Amount per liter 50 Amount per liter 50Molarity each ingredient) ppm (ppm of each) ppm (ul/liter) Ratio BleachNaBr 40 Bleach NaBr 40 Bleach NaBr 40 1:1 72 28 36 14 288 35 2:1 84 1642 8 336 20

TABLE 3 Dilutions for 100 ppm Cl₂ Equivalent Percentage (% of Amount perliter 100 Amount per liter 100 Molarity each ingredient) ppm (ppm ofeach) ppm (ul/liter) Ratio Bleach NaBr 40 Bleach NaBr 40 Bleach NaBr 401:1 72 28 72 28 576 70 2:1 84 16 84 16 672 40

TABLE 4 Dilutions for 150 ppm Cl₂ Equivalent Percentage (% of Amount perliter 150 Amount per liter 150 Molarity each ingredient) ppm (ppm ofeach) ppm (ul/liter) Ratio Bleach NaBr 40 Bleach NaBr 40 Bleach NaBr 401:1 72 28 108 42 864 105 2:1 84 16 126 24 1008 60NaBr = SANIBROM 40 Biocide (contains 40% sodium bromide, watersolution). Clorox ® bleach (Bleach) contains 12.5% chlorine.

Calculations for dilutions using the other biocides of this group thatwere tested were based on the following: Aquatize® biocide is a solutioncontaining 3.67% sodium chlorite, and Strabrm®909 biocide solution, itwas calculated as 1.57 times Cl₂ equivalent level. The results of thisgroup of tests are summarized in Tables 5-7. TABLE 5 Carcass BacteriaReduction Test Test Material Mean Bacteria Group Disinfectant LevelReduction¹ 1 Negative Control No reduction 2 Aquatize ® biocide(dilution 1:700) 4.25 × 10² 3 Aquatize ® biocide (dilution 1:350) 3.06 ×10³ 4 Aquatize ® biocide (dilution 1:250) 6.67 × 10³ 5 Clorox ® bleach,12.5% Cl₂ (dilution 1:2,500) 1.03 × 10² 6 Clorox ® bleach, 12.5% Cl₂(dilution 1:1,250) 5.11 × 10² 7 Clorox ® bleach, 12.5% Cl₂ (dilution1:800) 9.89 × 10² 8 SSBC (dilution 1:12,500) 2.41 × 10² 9 SSBC (dilution1:6,250) 5.87 × 10³ 10 SSBC (dilution 1:4,000) 4.69 × 10⁴ 11 Bleach andsodium bromide (1:1 mole ratio 3.52 × 10⁴ mix) Bleach 1:3,500 & NaBrdilution 1:28,000) 12 Bleach and sodium bromide (1:1 mole ratio 8.87 ×10⁴ mix) Bleach 1:1,750 & NaBr dilution 1:14,000) 13 Bleach and sodiumbromide (1:1 mole ratio 2.27 × 10⁵ mix) Bleach 1:1,200 & NaBr dilution1:9,300) 14 Bleach and sodium bromide (2:1 mole ratio 1.09 × 10³ mix)Bleach 1:3,000 & NaBr dilution 1:50,000) 15 Bleach and sodium bromide(2:1 mole ratio 1.55 × 10⁴ mix) Bleach 1:1,500 & NaBr dilution 1:25,000)16 Bleach and sodium bromide (2:1 mole ratio 5.21 × 10⁴ mix) Bleach1:1,000 & NaBr dilution 1:16,600) 17 Sodium bromide, 40% solution, 92Dilution 1:8,000 18 Sodium bromide, 40% solution, 6.54 × 10² Dilution1:4,000 19 Sodium bromide, 40% solution, 1.73 × 10³ Dilution 1:2,670¹The value represents an average of 10 birds per treatment.²Test group 1 carcass contained 2.67 × 10⁵ total bacteria count.

TABLE 6 Carcass Bacteria Reduction Results (% reduction) Test MeanBacteria Mean Bacteria Percent Bacteria Group Reduction¹ ReductionCount¹ Reduction From Control¹ 1 No reduction² ²) 267,000 Control 2 4.25× 10² 425 0.159% 3 3.06 × 10³ 3,060 1.146% 4 6.67 × 10³ 6,670 2.498% 51.03 × 10² 103 0.039% 6 5.11 × 10² 511 0.191% 7 9.89 × 10² 989 0.370% 82.41 × 10² 241 0.090% 9 5.87 × 10³ 5,870 2.199% 10 4.69 × 10⁴ 46,90017.566% 11 3.52 × 10⁴ 35,200 13.184% 12 8.87 × 10⁴ 88,700 33.221% 132.27 × 10⁵ 227,000 85.019% 14 1.09 × 10³ 1,090 0.408% 15 1.55 × 10⁴15,500 5.805% 16 5.21 × 10⁴ 52,100 19.513% 17 92 92 0.034% 18 6.54 × 10²654 0.245% 19 1.73 × 10³ 1,730 0.648%¹The value represents an average of 10 birds per treatment.²Test group 1 carcass contains 2.67 × 10⁵ total bacteria count.

TABLE 7 Chill Water Bacteria Count Test Test Material Mean BacteriaGroup Disinfectant Level Reduction¹ 1 Negative Control 5.11 × 10³ 2Aquatize ® biocide (dilution 1:700) 2.43 × 10³ 3 Aquatize ® biocide(dilution 1:350) 8.54 × 10² 4 Aquatize ® biocide (dilution 1:250) 2.21 ×10² 5 Clorox ® bleach, 12.5% Cl₂ (dilution 1:2,500) 5.43 × 10³ 6Clorox ® bleach, 12.5% Cl₂ (dilution 1:1,250) 4.24 × 10³ 7 Clorox ®bleach, 12.5% Cl₂ (dilution 1:800) 1.05 × 10³ 8 SSBC (dilution 1:12,500)4.83 × 10³ 9 SSBC (dilution 1:6,250) 1.64 × 10³ 10 SSBC (dilution1:4,000) 3.02 × 10² 11 Bleach and sodium bromide (1:1 mole ratio 2.55 ×10² mix) Bleach 1:3,500 & NaBr dilution 1:28,000) 12 Bleach and sodiumbromide (1:1 mole ratio 1.36 × 10² mix) Bleach 1:1,750 & NaBr dilution1:14,000) 13 Bleach and sodium bromide (1:1 mole ratio 43 mix) Bleach1:1,200 & NaBr dilution 1:9,300) 14 Bleach and sodium bromide (2:1 moleratio 1.98 × 10³ mix) Bleach 1:3,000 & NaBr dilution 1:50,000) 15 Bleachand sodium bromide (2:1 mole ratio 6.46 × 10² mix) Bleach 1:1,500 & NaBrdilution 1:25,000) 16 Bleach and sodium bromide (2:1 mole ratio 3.47 ×10² mix) Bleach 1:1,000 & NaBr dilution 1:16,600) 17 Sodium bromide (40%solution), 4.67 × 10³ Dilution 1:8,000 18 Sodium bromide (40% solution),3.49 × 10³ Dilution 1:4,000 19 Sodium bromide (40% solution), 2.23 × 10³Dilution 1:2,670¹The value represents bacteria count per mL of treatment water.

EXAMPLE 2

The procedure of Example 1 was repeated except that the materials testedfor microbiocidal activity were (a) sodium hypochlorite (Clorox®bleach), (b) the combination of sodium bromide and sodium hypochlorite,and (c) 1,3-dibromo-5,5-dimethylhydantoin (DBDMH), 100 birds were usedin this group of tests, and the chill water was composed per liter of950 mL of water, 50 mL of blood, 10 g of ground abdominal fat, 10 g ofmeat particles, and 10 g of skin with fat.

The experimental design used in this group of tests is summarized inTable 8. TABLE 8 Active Test Ingredient Group or equivalent TestMaterial Disinfectant Level 1 None No disinfectant¹ 2 Chlorine Clorox ®bleach 12.5% Cl₂ (dilution 1:2,500), (50 ppm) Contains 50 ppm chlorine²3 Chlorine Clorox ® bleach 12.5% Cl₂ (dilution 1:1.250) (100 ppm)Contains 100 ppm chlorine 4 Chlorine Clorox ® bleach 12.5% Cl₂ (dilution1:800) (150 ppm) Contains 150 ppm chlorine 5 Chlorine Bleach and LiquidSodium Bromide (1:1 mole (50 ppm total) ratio mix) Bleach dilution1:3,500 & NaBr dilution 1:28,000 Contains 50 ppm chlorine equivalent(1:1 Cl₂ equivalent) 6 Chlorine Bleach and Liquid Sodium Bromide (1:1mole (100 ppm total) ratio mix) Bleach dilution 1:1,750 & NaBr dilution1:14,000 Contains 100 ppm chlorine equivalent (1:1 Cl₂ equivalent) 7Chlorine Bleach and Liquid Sodium Bromide (1:1 mole (150 ppm total)ratio mix) Bleach dilution 1:1,200 & NaBr dilution 1:9,300 Contains 150ppm chlorine equivalent (1:1 Cl₂ equivalent) 8 Chlorine DBDMH(equivalent to 50 ppm Cl₂ level)- (50 ppm total) 0.9 g per literContains 50 ppm chlorine equivalent (1:1 Cl₂ equivalent) 9 ChlorineDBDMH (equivalent to 100 ppm Cl₂ level)- (100 ppm) 1.7 g per literContains 100 ppm chlorine equivalent (1:1 Cl₂ equivalent) 10 ChlorineDBDMH (equivalent to 150 ppm Cl₂ level)- (150 ppm) 3.4 g per literContains 150 ppm chlorine equivalent (1:1 Cl₂ equivalent)¹Negative control contained contaminated (bacteria 2.67 × 10⁵ per mL)water.²Positive control is normal poultry industry practice of adding 50 ppmchlorine.

The microbiocidal solution of this invention was prepared in thefollowing manner:

-   1. To form a stock solution, 100 g of    1,3-dibromo-5,5-dimethylhydantoin (DBDMH) was stirred into 10 liters    (10,000 mL) of water for 20 minutes. After filtration, the resulting    clear solution contains 1300 mg per liter as Br₂. This corresponds    to 580 mg per liter (or 580 ppm Cl₂) when expressed as Cl₂.-   2. The washing solution of DBDMH having a content of 50 ppm of Cl₂    equivalent solution was formed by mixing 875 mL of the above stock    solution with 10 liters (10,000 mL) of the above prepared chicken    chill water solution. The washing solutions of DBDMH containing 100    ppm Cl₂ equivalent and 150 ppm Cl₂ equivalent were prepared in the    same manner except that 1750 mL and 2625 mL, respectively, of the    above stock solution were mixed with separate 10-liter portions of    the above prepared chicken chill water solution.

Table 9 summarizes the results obtained in this group of tests. TABLE 9Carcass Bacteria Reduction Whole Bird Mean Chill Test Test MaterialBacteria Water Bacteria Group Disinfectant Level Reduction (%) Reduction(%)¹ 1 No disinfectant Control² Control 2 Clorox ® bleach³, 6.6% 8.2% 50ppm Cl₂ 3 Clorox ® bleach, 28.2% 32.8% 100 pp, Cl₂ 4 Clorox ® bleach41.1% 59.3% 150 ppm Cl₂ 5 NaBr 50 ppm Cl₂ 14.8% 18.4% equivalent +Bleach 6 NaBr 100 ppm Cl₂ 38.5% 41.6% equivalent + Bleach 7 NaBr 150 ppmCl₂ 73.5% 84.7% equivalent + Bleach 8 DBDMH, 50 ppm Cl₂ 99.9999%99.9999% equivalent 9 DBDMH, 100 ppm Cl₂ 99.9999% 99.9999% equivalent 10DBDMH, 150 ppm Cl₂ 99.9999% 99.9999% equivalent¹The value represents bacteria count per mL of treatment water.²Negative control contained contaminated (bacteria 2.67 × 10⁵ per mL)water.³Positive control is normal poultry industry practice of adding 50 ppmchlorine.

EXAMPLE 3

This group of tests was conducted to determine the effect of Clorox®bleach, Aquatize® biocide, and 1,3-dibromo-5,5-dimethylhydantoin (DBDMH)on carcass bacteria (Escherichia coli field strain) residual after1,5-hour in a chill tank “soup”. Tests were conducted with soups at pH7, pH 8 and pH 9 (adjusted by trisodium phosphate) for whole birdbacteria counts. Tests at pH 8 were conducted for individual bacteriacounts.

In general the tests involved normal processing of 56-day-old birds andimmersing the carcasses first in a warm bath containing 10⁴ per mLEscherichia coli, 10⁴ per mL Salmanella enteritidis, 10⁴ per mLPseudomonas aeruginosa, 10⁴ per mL Campylobacter jejuni, and 10⁴ per mLspoilage bacteria each from three strains (Listeria monocytogenes andShigella sonnei). The carcasses were then immersed in a chill tank“soup”, containing normal organic fluids (blood, fat, skin, and meatparticles) and containing the microbiocides on the test.

Tables 10 and 11 summarize the experimental design of these group oftests. TABLE 10 Whole Bird Bacteria Counts at pH 7, pH 8, and pH 9 TestGroup Test Material (Chill Tank) Reps Birds/Rep 1 None (Control) 5 10 2Clorox ® Bleach (20 ppm Cl₂ equivalent) 5 10 3 Aquatize ® (1:500dilution) 5 10 4 Aquatize ® (1:1000 dilution) 5 10 5 DBDMH (10 ppm Cl₂equivalent) 5 10 6 DBDMH (20 ppm Cl₂ equivalent) 5 10

TABLE 11 Individual Bird Bacteria Counts at pH 8 Test Group TestMaterial (Chill Tank) Reps Birds/Rep 7 None (Control) 5 5 8 Clorox ®Bleach (20 ppm Cl₂ equivalent) 5 5 9 DBDMH (10 ppm Cl₂ equivalent) 5 510 DBDMH (20 ppm Cl₂ equivalent) 5 5

The bacteria stock solution used for this group of tests was prepared bygrowing each bacteria sample in the appropriate broth shown in Table 12.Each such broth had a volume of at least 500 mL and the bacteria wereallowed to grow for at least 6 hours. The containers were observed andnot allowed to develop a heavy, cloudy visual appearance which wouldindicate that the growth had developed for too long a period. Thus thesolutions had the appearance of only being foggy or somewhat unclear.TABLE 12 Broth Treatments Organism¹ Broth Plating Media S. sonneiNutrient Broth Nutrient Agar L. monocytogenes Brain Heart Infusion BrainHeart Infusion Broth Agar E. coli Brain Heart Infusion Brain HeartInfusion Broth Agar S. enteritidis Tryptic Soy Broth Tryptic Soy Agar P.aeruginosa Tryptic Soy Broth Tryptic Soy Agar C. jejuni Brucella BrothBrucella Agar¹ Shigella sonnei, Listeria monocytogenes, Escherichia coli, Salmonellaenteritidis, Pseudomonas aeruginosa, and Campylobacter jejuni.

The microbiocidal solution of this invention was prepared in thefollowing manner:

-   1. To form a stock solution, 100 g of    1,3-dibromo-5,5-dimethylhydantoin (DBDMH) was stirred into 10 liters    (10,000 mL) of water for 20 minutes. After filtration, the resulting    clear solution contains 1300 mg per liter as Br₂. This corresponds    to 580 mg per liter (or 580 ppm Cl₂) when expressed as Cl₂.-   2. The chill water solution of DBDMH having a content of 10 ppm of    Cl₂ equivalent was formed by mixing 175 mL of the above stock    solution with 10 liters (10,000 mL) of the above prepared chicken    chill water solution. The chill water solution of DBDMH containing    20 ppm Cl₂ equivalent and 150 ppm Cl₂ equivalent were prepared in    the same manner except that 350 mL of the above stock solution were    mixed with another 10-liter portion of the above prepared chicken    chill water solution.

Table 13 shows the composition of the “chicken soup” used in thesetests. TABLE 13 Composition of “Chicken Soup” Material¹ Material per2100 mL² Water Added 1840 mL Bacteria Stock Solution 200 mL Blood 40 mLChicken Abdominal Fat (ground) 30 g Thigh Meat Particles 30 g Chickenskin with fat 10 g TOTAL 2100 mL equivalent¹The combined material was chilled overnight.²The material was ground and aggressively stirred prior to use.

The procedure used for whole bird wash sampling was as follows:

-   1. All samples were kept at <50 degrees Fahrenheit following    collection.-   2. Microbiological analyses of samples began within 24 hours of    sample collection.-   3. Information on the individual sample identification, date of    collection, time of collection (phase during shift), treatment group    and location of sample point were recorded on each sample bottle.-   4. At each defined sample time, carcasses were taken individually    from the processing line wearing latex or rubber gloves. The gloves    were rinsed with alcohol between each collection.-   5. Any excess fluid was drained off from the carcass. Each    individual carcass was transferred to a sterile stomacher bag.-   6. To each carcass contained in the sterile stomacher bag, 400 mL of    Butterfield's Phosphate Diluent (BPD) was added while making sure to    pour the BPD into the inside of the carcass cavity. The carcass was    rinsed inside and out with a rocking motion for one minute (ca. 35    RPM). This was best done by grasping the broiler carcass with one    hand and the closed top of the bag with the other then rocking with    a reciprocal motion in a 18-24 inch arc, assuring that all surfaces    (interior and exterior of the carcass) were rinsed.-   7. The rinse solutions from each stomacher bag was transferred into    the sample bottles, taking care to ensure that the information on    the date of collection, time of collection (phase during shift),    treatment group and location of sampler point matched that of the    sample.-   8. Each bottle was sealed with parafilm and placed into a styrene    container with crushed or dry ice or frozen freezer packs for    overnight delivery to a testing laboratory.-   9. All filled styrene containers were held in a chilled (not below    freezing) area until within 1 to 2 hours of courier collection for    shipment.

Quantitative or qualitative determinations for bacterial organisms wereconducted according to the following methodologies:

Aerobic plate counts—Counting rules according to BAM 8th ed., Chapter 3.

Coliform and E. coli counts—AOAC, 991.14, Petrifilm.

Salmonella—AOAC 986.35, ELISA presumptive screen.

Salmonella—USDA LC-75, incidence.

Campylobacter—USDA LC-69, incidence.

Listeria—USDA LC-57, incidence.

In greater detail the trial events and experimental design used in thisgroup of tests were as follows:

-   a) Test microorganisms used were:    -   Escherichia coli ATCC 11229    -   Pseudomonas aeruginosa ATCC 15442    -   Salmonella enteritidis ATCC 13076    -   Shigella sonnei ATCC 9290    -   Listeria monocytogenes ATCC 7644    -   Campylobacter jejuni ATCC 29428-   b) Test Procedure: All test strains were grown individually at    35° C. for 24 hours in the media specified in Table 12. Cells were    harvested by centrifugation at 10,000×g for 10 minutes and washed    twice with Butterfield's Phosphate Buffer (BPB of pH 7.2). Cells    were resuspended in BPB to obtain a cell suspension of approximately    1.0×10⁸ CFU/mL for each microorganism. The target inoculum levels    were approximately 10⁶ CFU/mL in the final test solutions. In the    cases of S. enteritidis and P. aeruginosa the species were washed by    pouring into prepared sterile centrifuge tubes with cheesecloth    filters. The culture was then pelleted and washed using above    techniques and repeated 3 times.-   c) The birds (56 days old) were processed under normal commercial    conditions.-   d) The bacteria were added to a large batch of the “chicken soup”,    and then aliquots of the resultant mixture were distributed equally    among the chill waters used for each test. Then the particular    disinfectant composition under test was added to one of the chill    waters. The chill waters each contained 10⁴ per mL Escherichia coli,    10⁴ per mL Salmonella enteritidis, 10⁴ per mL Campylobacter jejuni,    and 10⁴ per mL spoilage bacteria each from three strains (Listeria    monocytogenes, Pseudomonas aeruginosa, and Shigella sonnei).-   e) Birds were added to each of ten 50-gallon containers containing    these respective treatments (or control) and were kept in the    containers for the 1.5 hour chilling period.-   f) During the 1.5 hour chilling period, the contents were vigorously    stirred every 10 minutes.-   g) After the 1.5 hour chilling period, the whole birds were placed    in individual sterile stomacher bags and the whole bird rinse (as    described above) was conducted and samples of the rinse were placed    on the appropriate agar plates. The plates were placed in the    incubator for 24 hours at 37° C. Then the plates were read after 24    hours to determine total count on each plate.

The results of this group of tests are summarized in Tables 14 and 15.TABLE 14 Whole Bird Total Aerobic Bacteria (% Reduction)¹ WaterTreatment Water pH 7 Water pH 8 Water pH 9 None (Control) — — — Clorox ®Bleach 15% 15%  2% (20 ppm Cl₂ equivalent) Aquatize ® biocide 76% 71%64% (1:500 dilution) Aquatize ® biocide 42% 45% 33% (1:1000 dilution)DBDMH 85% 82% 78% (10 ppm Cl₂ equivalent) DBDMH 99% 98% 96% (20 ppm Cl₂equivalent)¹Each value represents 50 birds per treatment.

TABLE 15 Disinfecting Treatment (average bacteria count per bird)^(2,3)Clorox Bleach DBDMH DBDMH Organism¹ Control (20 ppm) (10 ppm) (20 ppm)S. sonnei 4551 3552 456 12 L. monocytogenes 2463 2065 262 6 E. coli 30552759 357 4 S. enteritidis 3969 3160 560 10 P. aeruginosa 2783 2280 289 9C. jejuni 1282 981 183 15 Mean % Reduction — 18.3% 85.8% 98.8% FromControl¹ Escherichia coli, Salmonella enteritidis, Pseudomonas aeruginosa,Campylobacter jejuni, listeria monocytogenes, and Shigella sonnei²NOTE:Cross contamination is more likely in a processing environment wherebirds were processed and samples taken for individual culturedetermination.³Each value represents 25 birds per treatment.

EXAMPLE 4

A study was conducted to determine the effect of Clorox® bleach, and1,3-dibromo-5,5-dimethylhydantoin (DBDMH) on carcass bacteria residualafter 1.5 hour in a chill tank solution and spoilage 20-day shelf lifelongevity (caused by bacteria contamination). Tests were conducted at pH8 (adjusted by trisodium phosphate). Skin pigmentation (Minolta ColorMeter L value or Lightness, a value or redness and b value oryellowness) were determined before and post-processing.

In general the study involved normal processing of 56-day-old birds,immersing carcasses first in a warm bath containing 10⁴ per mLEscherichia coli, 10⁴ per mL Salmonella enteritidis, 10⁴ per mLPseudomonas aeruginosa, 10⁴ per mL Campylobacter jejuni, and 10⁴ per mLspoilage bacteria each from three strains (Listeria monocytogenes andShigella sonnel). Carcass were then immersed in a chill tank “soup”,containing normal organic fluids (blood, fat, skin, and meat particles)and containing various disinfectants (termed test materials).

Four test groups of birds were tested at pH 8 for whole bird bacteriacounts. Table 16 sets forth the experimental design for these wholebacteria count tests. TABLE 16 Test Group Test Material (Chill Tank)Reps Birds/Rep 1 None (Control) 6 10 2 Clorox ® bleach (20 ppm Cl₂equivalent) 6 10 3 DBDMH (10 ppm Cl₂ equivalent) 6 10 4 DBDMH (20 ppmCl₂ equivalent) 6 10

A DBDMH stock solution and test solutions, a bacteria stock solution,and a “chicken soup” were prepared as in Example 3. In addition, thebacterial broth treatments, the whole bird wash sampling procedure, andthe methodologies used for quantitative or qualitative determinationsfor bacterial organisms were conducted as in Example 3.

In greater detail the trial events and experimental design used in thisgroup of tests were as follows:

-   a) Test microorganisms used were:    -   Escherichia coli ATCC 11229    -   Pseudomonas aeruginosa ATCC 15442    -   Salmonella enteritidis ATCC 13076    -   Shigella sonnei ATCC 9290    -   Listeria monocytogenes ATCC 7644    -   Campylobacter jejuni ATCC 29428-   b) Test Procedure: All test strains were grown individually at    35° C. for 24 hours in the media specified in Table 12. Cells were    harvested by centrifugation at 10,000×g for 10 minutes and washed    twice with Butterfield's Phosphate Buffer (BPB of pH 7.2). Cells    were resuspended in BPB to obtain a cell suspension of approximately    1.0×10⁸ CFU/mL for each microorganism. The target inoculum levels    were approximately 10⁶ CFU/mL in the final test solutions. In the    cases of S. enteritidis and P. aeruginosa the species were washed by    pouring into prepared sterile centrifuge tubes with cheesecloth    filters. The culture was then pelleted and washed using above    techniques and repeated 3 times.-   c) The birds (56 days old) were processed under normal commercial    conditions.-   d) The bacteria were added to a large batch of the “chicken soup”,    and then aliquots of the resultant mixture were distributed equally    among the chill waters used for each test. Then the particular    disinfectant composition under test was added to one of the chill    waters. The chill waters each contained 10⁴ per mL Escherichia coli,    10⁴ per mL Salmonella enteritidis, 10⁴ per mL Campylobacter jejuni,    and 10⁴ per mL spoilage bacteria each from three strains (Listeria    monocytogenes, Pseudomonas aeruginosa, and Shigella sonnei).-   e) Birds were added to each of ten 50-gallon containers containing    these respective treatments (or control) and were kept in the    containers for the 1.5 hour chilling period.-   f) During the 1.5 hour chilling period, the contents were vigorously    stirred every 10 minutes.-   g) After the 1.5 hour chilling period, the whole birds were placed    in a commercial refrigerator for 20-days of storage.-   h) Skin pigmentation (using Minolta Color Meter L or Lightness, a or    redness and b or yellowness) were determined on all birds before and    immediately after post-processing chilling.-   i) For Day 0, a total of 5 whole birds per treatment were randomly    chosen from each treatment and placed in individual sterile    stomacher bag and the whole bird rinse (as described in Example 3)    was carried out and samples of the rinse were placed on appropriate    agar plates.-   j) For each of succeeding days 2, 4, 6, 8, 10, 12, 14, 16, 18, and    20, a total of 5 whole birds per treatment were randomly chosen from    each treatment and placed in individual sterile stomacher bags and    the whole bird rinse (as described in Example 3) was conducted and    samples of the rinse were placed on the appropriate agar plates.-   k) All of the treated agar plates were placed in an incubator for 24    hours at 35° C. Plates were read after 24 hours to determine total    count on each plate.

The results of these tests are summarized in Tables 17-30. TABLE 17Percentage of Total Bacteria Reduction From Control (Dayspost-processing) Water Day Day Day Day Day Day Day Day Day Day DayTreatment 0 2 4 6 8 10 12 14 16 18 20 None — — — — — — — — — — —(Control) Clorox ® 22.5 23.1 22.2 25.2 26.0 25.7 25.9 26.5 23.2 23.1220.5 Bleach (20 ppm) DBDMH 77.8 77.3 76.8 77.1 74.6 71.9 69.2 66.2 61.958.5 53.7 (10 ppm) DBDMH 99.5 99.4 99.2 98.5 97.3 95.1 91.2 84.3 71.268.0 67.2 (20 ppm)

TABLE 18 Average skin TBA Values¹ (Days post-processing) Water Day DayDay Day Day Day Day Day Day Day Day Treatment 0 2 4 6 8 10 12 14 16 1820 None 0.14a 0.31a 0.45a 0.69a 0.88a 1.23a 1.36a 1.66a 2.08a 2.39a3.02a (Control) Clorox ® 0.10a 0.42a 0.68a 0.72a 0.90a 1.10a 1.49a 1.73a2.19a 2.51a 2.88a Bleach (20 ppm) DBDMH 0.20a 0.54a 0.79a 0.54a 0.76a1.20a 1.77a 1.94a 2.33a 2.45a 2.92a (10 ppm) DBDMH 0.22a 0.36a 0.46a0.71a 0.75a 1.22a 1.53a 1.87a 2.19a 2.68a 2.73a (20 ppm)NOTE:¹Means within a row without a common superscript are significantlydifferent (P < 0.05) as determined by Least Significant Difference.

TABLE 19 Skin Pigmentation Value (Minolta Color Meter)¹ Mean Pre-ChillMean Post-Chill Minolta Value Minolta Value Water Treatment L a b L a bNone (Control) 62.84a 5.32a 15.42a 58.84a 5.93a 16.84a Clorox ® 63.62a5.49a 15.94a 58.84a 5.64a 16.16a Bleach (20 ppm) DBDMH (10 ppm) 61.55a5.14a 15.63a 58.84a 6.09a 16.22a DBDMH (20 ppm) 60.77a 5.69a 15.67a58.84a 6.24a 16.37aNOTE:¹Means within a row without a common superscript are significantlydifferent (P < 0.05) as determined by Least Significant Difference.

TABLE 20 Effect of Disinfection Treatment on Day 0 DisinfectingTreatment (average bacteria count per bird) Clorox bleach DBDMH DBDMHOrganism Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 3687 2948 845 8 L.monocytogenes 2569 2281 528 13 E. coli 3879 2310 861 22 S. enteritidis1678 1064 292 12 P. aeruginosa 2974 2681 743 6 C. jejuni 2276 1935 51917 Mean % Reduction — 22.5% 77.8% 99.5% From Control

TABLE 21 Effect of Disinfection Treatment on Day 2 DisinfectingTreatment (average bacteria count per bird) Clorox DBDMH DBDMH OrganismControl (20 ppm) (10 ppm) (20 ppm) S. sonnei 4119 3241 962 12 L.monocytogenes 2749 2442 601 19 E. coli 4193 2604 966 31 S. enteritidis1921 1191 344 18 P. aeruginosa 3313 2889 820 9 C. jejuni 2534 2114 57325 Mean % Reduction — 23.1% 77.3% 99.4% From Control

TABLE 22 Effect of Disinfection Treatment on Day 4 DisinfectingTreatment (average bacteria count per bird)² Clorox DBDMH DBDMHOrganism¹ Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 4664 3528 1101 19L. monocytogenes 2920 2751 670 28 E. coli 4379 3001 1050 49 S.enteritidis 2152 1309 394 27 P. aeruginosa 3592 3127 931 13 C. jejuni2830 2267 627 39 Mean % Reduction — 22.2% 76.8% 99.2% From Control

TABLE 23 Effect of Disinfection Treatment on Day 6 DisinfectingTreatment (average bacteria count per bird)² Clorox DBDMH DBDMHOrganism¹ Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 5424 3802 1288 37L. monocytogenes 3176 3071 741 55 E. coli 4769 3142 1124 100 S.enteritidis 2426 1347 433 55 P. aeruginosa 4141 3454 1013 25 C. jejuni3113 2423 671 78 Mean % Reduction — 25.2% 77.1% 98.5% From Control

TABLE 24 Effect of Disinfection Treatment on Day 8 DisinfectingTreatment (average bacteria count per bird)² Clorox DBDMH DBDMHOrganism¹ Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 5969 4008 1604 76L. monocytogenes 3407 3474 880 107 E. coli 5194 3438 1364 204 S.enteritidis 2764 1519 507 104 P. aeruginosa 4768 3798 1268 48 C. jejuni3353 2594 834 157 Mean % Reduction — 26.0% 74.6% 97.3% From Control

TABLE 25 Effect of Disinfection Treatment on Day 10 DisinfectingTreatment (average bacteria count per bird)² Clorox DBDMH DBDMHOrganism¹ Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 6292 4415 1954156 L. monocytogenes 3854 3767 1096 218 E. coli 5683 3694 1621 401 S.enteritidis 3116 1605 616 212 P. aeruginosa 5243 4305 1485 91 C. jejuni3589 2844 1043 294 Mean % Reduction — 25.7% 71.9% 95.1% From Control

TABLE 26 Effect of Disinfection Treatment on Day 12 DisinfectingTreatment (average bacteria count per bird)² Clorox DBDMH DBDMHOrganism¹ Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 6890 5030 2347323 L. monocytogenes 4348 4195 1335 442 E. coli 6316 3902 2063 775 S.enteritidis 3461 1819 740 413 P. aeruginosa 5743 4720 1730 186 C. jejuni4133 3213 1309 594 Mean % Reduction — 25.9% 69.2% 91.2% From Control

TABLE 27 Effect of Disinfection Treatment on Day 14 DisinfectingTreatment (average bacteria count per bird)² Clorox DBDMH DBDMHOrganism¹ Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 7768 5313 2848657 L. monocytogenes 4781 4755 1564 843 E. coli 6762 4279 2581 1453 S.enteritidis 3901 2055 919 832 P. aeruginosa 6426 5200 2055 363 C. jejuni4454 3446 1551 1191 Mean % Reduction — 26.5% 66.2% 84.3% From Control

TABLE 28 Effect of Disinfection Treatment on Day 16 DisinfectingTreatment (average bacteria count per bird) ² Clorox DBDMH DBDMHOrganism¹ Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 7970 6108 35131286 L. monocytogenes 5263 5228 1901 1646 E. coli 7201 4692 3005 2933 S.enteritidis 4281 2328 1081 1711 P. aeruginosa 6969 6005 2560 700 C.jejuni 4898 3733 1880 2259 Mean % Reduction — 23.2% 61.9% 71.2% FromControl

TABLE 29 Effect of Disinfection Treatment on Day 18 DisinfectingTreatment (average bacteria count per bird)^(2,3) Clorox DBDMH DBDMHOrganism¹ Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 9004 6957 42421604 L. monocytogenes 5799 5694 2221 1985 E. coli 7725 5097 3617 3645 S.enteritidis 4835 2613 1286 2074 P. aeruginosa 7814 6869 3087 826 C.jejuni 5319 3900 2359 2835 Mean % Reduction — 23.1% 58.5% 68.0% FromControl

TABLE 30 Effect of Disinfection Treatment on Day 20 DisinfectingTreatment (average bacteria count per bird)^(2,3) Clorox DBDMH DBDMHOrganism¹ Control (20 ppm) (10 ppm) (20 ppm) S. sonnei 9288 7409 49411834 L. monocytogenes 6419 6506 2678 2238 E. coli 8272 5635 4460 4036 S.enteritidis 5335 2976 1513 2258 P. aeruginosa 8604 7886 3853 908 C.jejuni 5789 4332 2789 3059 Mean % Reduction — 20.5% 53.7% 67.2% FromControl

In Tables 19-30 each figure on average bacteria count per birdrepresents the average of 5 birds.

EXAMPLE 5

The objective of this study was to determine the effect of bleachmicrobiocidal control (20 ppm Cl₂ equivalent) and of microbiocidalcontrol with 1,3-dibromo-5,5-dimethyl-hydantoin (DBDMH) on organoleptictaste evaluation of both breast and thigh meat. Formal trained tastepanel evaluation was conducted. The trial was conducted using 49-day oldbirds which were processed unchallenged with external sources ofbacteria and under sterile conditions.

A total of 120 birds were used in this study. Sixty of the birds servedas a control group. These were subjected to treatment in a chill tankcontaining Clorox® bleach at a 20 ppm Cl₂ equivalent level. The other 60birds were treated in a chill tank in the same fashion except that thechilling water contained DBDMH at the level of 20 ppm Cl₂ equivalent.During the 1.5 hour chilling period in the chill tank, the contents ofthe tank were vigorously stirred every 10 minutes. After the 1.5 hourchilling period, the whole birds were individually bagged and placed ina commercial refrigerator for 20 days of storage. After aging,individual breast and thigh samples were cut and cooked to an internaltemperature of 190° F. Taste evaluation was determined using 10 trainedtaste panel experts. A Ranking System (“1” or “2”) was used where “1”represents the better tasting sample. A simple average of subjectevaluations or rankings per person were used. Statistical evaluation wasemployed by using each subject as a block employed delta 0.05.

Tables 31 and 32 set forth the results of these taste evaluations. TABLE31 Effect of Chill Tank Water Treatment On Taste Preference (Breast MeatEvaluation) SUMMARY - Tasting Ranking¹ Water Treatment S1 S2 S3 S4 S5 S6S7 S8 S9 S10 Mean² None (20 ppm Cl₂ 2 1 1 1 2 1 1 2 1 2 1.4 a equivalentbleach control) DBDMH (20 ppm Cl₂ equivalent) 1 2 2 2 1 2 2 1 2 1 1.6 a¹S(subject) = trained taste panelist subject number.NOTE:²Means within a row without a common superscript are significantlydifferent (P < 0.05) as determined by Least Significant Difference.

TABLE 32 Effect of Chill Tank Water Treatment On Taste Preference (ThighMeat Evaluation) SUMMARY - Tasting Ranking¹ Water Treatment S1 S2 S3 S4S5 S6 S7 S8 S9 S10 Mean² None (20 ppm Cl₂ equivalent 1 2 2 1 1 2 2 2 1 21.6 a bleach control) DBDMH (20 ppm Cl₂ equivalent) 2 1 1 2 2 1 1 1 2 11.4 a¹S(subject) = trained taste panelist subject number.NOTE:²Means within a row without a common superscript are significantlydifferent (P < 0.05) as determined by Least Significant Difference.

EXAMPLE 6

The objective of this study was to determine the effect of Clorox®bleach and 1,3-dibromo-5,5-dimethylhydantoin (DBDMH) on individualcarcass bacteria field strains after 1.5 hour in a chill tank solutionand spoilage 20-day shelf life longevity (caused by bacteriacontamination) in a Graded Level Study Model. After normal processing of56-day-old birds, carcasses were immersed first in a warm bathcontaining 10⁴ CFU's per mL Escherichia coli, 10⁴ CFU's per mLSalmonella enteritidis, 10⁴ CFU's per mL Pseudomonas aeruginosa, 10⁴CFU's per mL Campylobacter jejuni, and 10⁴ CFU's per mL spoilagebacteria each from two strains (Listeria monocytogenes and Shigellasonnei). Carcasses were then immersed in a chill tank “soup”, containingnormal organic fluids (blood, fat, skin, and meat particles) andcontaining various disinfectants (termed test materials). These testswere conducted at pH 8 (adjusted by trisodium phosphate). Skinpigmentation (Minolta Color Meter L value or Lightness, a value orredness and b value or yellowness) was determined before and afterprocessing. Post-chilling skin bacteria of various strains weredetermined over a 20-day period. Sensory evaluation was determined todemonstrate spoilage times and shelf-life. After salmonella infection inchill tanks, USDA HACCP salmonella detection was simulated and reported.

The materials tested and the experimental design of these test were assummarized in Table 33. TABLE 33 Test Group Test Material (Chill Tank)Reps Birds/Rep 1 None (Control) 10 12 2 Clorox ® bleach (20 ppm Cl₂equivalent 10 12 3 DBDMH (5 ppm Cl₂ equivalent) 10 12 4 DBDMH (10 ppmCl₂ equivalent) 10 12 5 DBDMH (15 ppm Cl₂ equivalent) 10 12 6 DBDMH (20ppm Cl₂ equivalent) 10 12 7 DBDMH (25 ppm Cl₂ equivalent) 10 12

A DBDMH stock solution and DBDMH test solutions of the concentrationsspecified in Table 33, a bacteria stock solution, and a “chicken soup”were prepared as in Example 3. In addition, the bacterial brothtreatments, the whole bird wash sampling procedure, and themethodologies used for quantitative or qualitative determinations forbacterial organisms were conducted as in Example 3.

The trial events and experimental design used in this group of testswere the same as in Example 5 with the following exceptions:

-   a) The temperature during the 20-day period of storage in the    refrigerator was 4° F.-   b) Observations of the degree of “bloating” (defined as water or air    additions under the skin area considered objectionable) were    conducted on all processed birds.-   c) On each of sampling days 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, and    20, ten carcasses from each treatment were analyzed by removing 23.8    cm² of skin from the breast right up to the neck using a template    and a sterile scalpel. Each skin sample was placed in a bag with 15    mL Butterfield's Phosphate Buffer Solution (BPBS) added and treated    in a Stomacher bag for 60 seconds. A 10-fold dilution series of the    mixture was made in BPBS and two parallel samples of 20 mL each were    spread on the appropriate plate count agar for determination of the    total viable numbers. The plates were incubated at 35° C. for 24    hours. Mean values were calculated from the two determinations of    the three samples taken from each combination of chilling and    storage. Bacterial numbers were reported as pooled or averaged log₁₀    colony-forming units (CFU's) per square centimeter.-   d) Also on sampling day 0, 102 total of the remaining 110 carcasses    from each treatment (all bloating and oddly processed birds were    removed) were “whole bird” washed by the sampling procedure    described in Example 3. Salmonella detection were noted and reported    as number of positive salmonella colonies per 51 birds and % of    total.

Tables 34-37 summarize the results of this group of tests. TABLE 34Salmonella Positive Samples (Number per 51)¹ (Birds were inoculated withSalmonella Water Treatment prior to chilling) None (Control) 32/51(62.74%) Clorox ® Bleach 11/51 (22.57%) (20 ppm) DBDMH (5 ppm)  7/51(13.73%) DBDMH (10 ppm) 4/51 (7.84%) DBDMH (15 ppm) 2/51 (3.92%) DBDMH(20 ppm) 1/51 (1.96%) DBDMH (25 ppm) 0/51 (0.00%)¹Twelve (12) per 51 or less is considered to be statistically acceptableby USDA HACCP standards. A total of 102 birds were used to determinesalmonella positive samples and a simple average determined.

TABLE 35 Water Treatment Birds (Number per 60 birds processed)¹ None(Control) 1/120 (0.83%) Clorox ® Bleach (20 ppm) 0/120 (0.00%) DBDMH (5ppm) 2/120 (1.67%) DBDMH (10 ppm) 0/120 (0.00%) DBDMH (15 ppm) 1/120(0.83%) DBDMH (20 ppm) 0/120 (0.00%) DBDMH (25 ppm) 1/120 (0.83%)¹Four (4) or more per treatment is considered to be highlyobjectionable.

TABLE 36 Sensory Score (days post-processing)^(1,2) Water Treatment 5days 10 days 15 days 20 days None (Control) 5.6 c 7.3 c 8.2 c 9.0 dClorox ® bleach (20 ppm) 3.8 b 3.6 b 5.5 b 7.1 c DBDMH (5 ppm)  2.4 ab3.2 b 3.9 a 5.6 a DBDMH (10 ppm) 1.9 ab 2.3 a 3.4 a 4.8 a DBDMH (15 ppm)1.3 a 2.1 a 2.6 a 4.9 a DBDMH (20 ppm) 1.1 a 1.8 a 2.7 a 4.3 a DBDMH (25ppm) 1.4 a 2.1 a 2.3 a 4.6 a¹Continuous scale for non-structured fresh inside carcass odor sensoryattributes ranges from value 1.0 (the lowest intensity) to value 9.0(the highest intensity).NOTE:Means within a row without a common superscript are significantlydifferent (P < 0.05) as determined by Least Significant Difference.²Five (5) or more is considered to be highly objectionable.

TABLE 37 Skin Pigmentation¹ Mean Pre-Chill Mean Post-Chill MinoltaValue² Minolta Value² Water Treatment L a B L a b None (Control) 59.72 a4.34 a 13.67 a 51.84 a 5.12 a 15.27 a Clorox ® Bleach 60.76 a 4.93 a13.74 a 55.81 a 5.08 a 15.49 a (20 ppm) DBDMH (5 ppm) 58.80 a 4.67 a13.61 a 52.68 a 5.42 a 15.64 a DBDMH (10 ppm) 59.97 a 4.31 a 13.64 a53.19 a 5.69 a 15.75 a DBDMH (15 ppm) 58.43 a 4.84 a 13.81 a 54.21 a5.55 a 15.64 a DBDMH (20 ppm) 58.54 a 4.99 a 13.67 a 53.74 a 5.49 a15.80 a DBDMH (25 ppm) 58.97 a 4.68 a 13.50 a 54.25 a 5.63 a 15.76 aNOTE:¹Means within a row without a common superscript are significantlydifferent (P < 0.05) as determined by Least Significant Difference.²Skin pigmentation (Minolta Color Meter L value or Lightness, a value orredness and b value or yellowness).

Results from the above tests on the effect of chill tank treatment ongrowth of Pseudomonas species on the chicken skin are graphicallydepicted in FIG. 1. FIG. 2 depicts graphically the results of the abovetests on the effect of chill tank treatment on growth of total aerobicbacteria on the chicken skin.

EXAMPLE 7

A study was carried out to determine the effectiveness of severalmicrobiocidal compounds of this invention, as well as sodiumhypochlorite when used as carcass rinses. The microbiocides of thisinvention used in this study were 1,3-dibromo-5,5-dimethylhy-dantoin(DBDMH), N,N′-bromochloro-5,5-dimethylhydantoin (BCDMH) and Stabrom® 909biocide (Albemarle Corporation), a concentrated alkaline aqueoussolution produced from bromine chloride and sulfamate anion (SSBC).

After normal processing of 56-day-old birds, carcasses were immersedfirst in a warm bath containing 10⁴ per mL Escherichia coli, 10⁴ per mLSalmonella enteritidis, 10⁴ per mL Pseudomonas aeruginosa, 10⁴ per mLCampylobacter jejuni, and 10⁴ per mL spoilage bacteria each from twostrains (Listeria monocytogenes and Shigella sonnei). Carcasses werethen immersed in a chill tank “soup”, containing normal organic fluids(blood, fat, skin, and meat particles) and containing variousdisinfectants (termed test materials). These whole bird bacteria counttests were conducted at pH 8. The effect of the test compounds on skinpigmentation was determined by use of Minolta Color Meter L value orLightness, a value or redness and b value or yellowness. Post-chillingskin bacteria of various strains were determined over a 20-day period.Spoilage, using sensory odors as a model, determined time required tocreate a putrid/ammonia-like odor. After salmonella infection in chilltanks, USDA HACCP salmonella detection was simulated and reported. Table38 describes the test material dosages and overall design of this groupof tests. TABLE 38 Test Group Test Material (Chill Tank) Reps Birds/Rep1 None (Control) 10 12 2 Clorox ® bleach (20 ppm Cl₂ equivalent) 10 12during chilling 3 DBDMH (20 ppm Cl₂ equivalent) 10 12 during chilling 4BCDMH (20 ppm Cl₂ equivalent) 10 12 during chilling 5 SSBC carcass spray1 10 (3% liquid pre-chill application)

DBDMH and BCDMH stock solutions and diluted test solutions (20 ppm Cl₂equivalent), a bacteria stock solution, and a “chicken soup” wereprepared as in Example 3 except that the Stabrom® 909 biocideconcentrate was diluted by adding 30 mL per liter of water just prior toapplication. This diluted solution was sprayed on the birds, both insideand outside, in quantities of 200 mL per bird. In addition, thebacterial broth treatments, the whole bird wash sampling procedure, andthe methodologies used for quantitative or qualitative determinationsfor bacterial organisms were conducted as in Example 3.

The details concerning the trial events used as well as the detailedexperimental design used in these tests were the same as described inExample 6. The only exceptions were:

-   a) In the case of the birds of Test Group 5 (note Table 38), while    the carcass was still warm, the 10 birds were each sprayed both    internally and externally, using a misting hand-held nozzle, with    200 mL of the 3% solution of Stabrom 909 biocide (SSBC). Previous    quality control trials using dye had ensured that complete carcass    coverage was achieved with the use of 200 mL of liquid spray. The    spray was allowed to stay on the warm carcasses for 60 seconds.-   b) The treatment on sampling day 0 of 102 total of the remaining 110    carcasses from each treatment involving “whole bird” washing and    Salmonella detection, all as described in Example 6, was applied    only to the birds of Test Groups 1-4 (note Table 38).

Tables 39-42 summarize the results of this group of tests. The effect ofthe chill tank treatment of this Example on growth of Pseudomonasspecies on chicken skin are graphically depicted in FIG. 3. FIG. 4depicts graphically the results of the tests of this Example on theeffect of chill tank treatment on growth of total aerobic bacteria onthe chicken skin. TABLE 39 Salmonella positive samples (Number 51)¹(Birds were inoculated with Water Treatment Salmonella prior tochilling) None (Control) 21/51 (41.18%)  Clorox ® bleach (20 ppm) 8/51(15.68%) DBDMH (20 ppm Cl₂ equivalent) 1/51 (1.96%)  during chillingBCDMH (20 ppm Cl₂ equivalent) 6/51 (11.76%) during chilling¹Twelve (12) per 51 or less is considered to be statistically acceptableby USDA HACCP standards. A total of 102 birds were used to determinesalmonella positive samples and a simple average determined.

TABLE 40 Bloating Water Treatment (Number per 60 birds processed)¹ None(Control) 0/120 (0.00%) Clorox ® bleach (20 ppm) 0/120 (0.00%) DBDMH (20ppm Cl₂ equivalent) 0/120 (0.00%) during chilling BCDMH (20 ppm Cl₂equivalent) 0/120 (0.00%) during chilling¹Four (4) or more per treatment is considered to be highlyobjectionable.

TABLE 41 Sensory Score (days post-processing)^(1,2) Water Treatment 5days 10 days 15 days 20 days None (Control) 2.4 b 4.8 c 6.9 c 9.0 dClorox ® bleach (20 ppm) 1.3 ab 2.4 b 4.6 b 6.8 c DBDMH (20 ppm Cl₂equivalent) 0.6 a 1.2 a 3.2 a 3.4 a during chilling BCDMH (20 ppm Cl₂equivalent) 1.4 ab 1.8 ab 2.7 a 4.8 b during chilling¹Continuous scale for non-structured fresh inside carcass odor sensoryattributes ranges from value 1.0 (the lowest intensity) to value 9.0(the highest intensity).NOTE:Means within a row without a common superscript are significantlydifferent (P < 0.05) as determined by Least Significant Difference.²Five (5) or more is considered to be highly objectionable.

TABLE 42 Skin Pigmentation¹ Mean Pre-Chill Mean Post-Chill MinoltaValue² Minolta Value² Water Treatment L a b L a b None (Control) 52.61 a3.25 a 11.43 a 47.21 a 4.24 a 12.44 a Clorox ® bleach 52.76 a 3.32 a11.84 a 47.43 a 4.85 a 12.67 a (20 ppm) DBDMH (20 ppm 52.23 a 3.13 a11.63 a 48.02 a 4.69 a 12.47 a Cl₂ equivalent) during chilling BCDMH (20ppm 52.11 a 3. 82 a 11.26 a 46.93 a 4.44 a 12.60 a Cl₂ equivalent)during chilling SSBC Carcass Spray 52.61 a 3.67 a 11.15 a 47.03 a 4.51 a12.55 a (3% liquid pre- chill application)NOTE:¹Means within a row without a common superscript are significantlydifferent (P < 0.05) as determined by Least Significant Difference.²Skin pigmentation (Minolta Color Meter L value or Lightness, a value orredness and b value or yellowness)³All treatment skin pigmentation were measured on 120 birds, except forSSBC where only 10 birds were employed.

A number of tests have been carried out demonstrating the microbiocidaleffectiveness of several microbiocides in eradicating or controllingvarious bacteria species of the types present in poultry processingsystems.

One such series of tests involved determinations of microbiologicalcontrol against Escherichia coli bacteria. Another set of tests involveddeterminations of microbiological control against Enterococcus faecium.In each case, comparative tests were carried out in the same mannerutilizing the AOAC test method. Such test involves exposing a culture ofthe microorganism to various concentrations of a test solution preparedfrom an aqueous stock solution of the compound under test. At varioustime intervals the halogen in the test suspensions is chemicallyneutralized, and the amount of viable bacteria remaining is enumeratedby plating onto nutrient agar and incubating for 2 days at 37° C.Results are expressed at the log₁₀ colony forming units (CFU). Theconcentration of the compound required to achieve complete kill (i.e.,no viable bacteria remain) within 30 seconds is determined in the test.

Table 43 summarizes the data obtained in the tests using respectively,1,3-dibromo-5,5-dimethylhydantoin (DBDMH) andN,N′-bromochloro-5,5-dimethylhydantoin (BCDMH) and in which themicroorganism in each case was Escherichia coli. It can be seen that1,3-dibromo-5,5-dimethylhydantoin passed the test at one milligram ofbromine, as Br₂₁ per liter of water, as evidenced by the complete killwithin 30 seconds, whereas 1,3-bromochloro-5,5-dimethylhydantoinrequired two milligrams of bromine, as Br₂, per liter of water toachieve complete kill within 30 seconds. TABLE 43 EFFECTIVENESS AGAINSTESCHERICHIA COLI Log₁₀ CFU Log₁₀ CFU Concentration Recovered UsingRecovered Using mg/L as Br₂ Contact Time DBDMH BCDMH 0.5 mg/L 30sec >4.48 >4.48 1 min 1.70 4.46 2 min 0 1.65 3 min 0 0 4 min 0 0 5 min 00 10 min 0 0 1.0 mg/L 30 sec 0 >4.48 1 min 0 0.7 2 min 0 0 3 min 0 0 4min 0 0 5 min 0 0 10 min 0 0 2.0 mg/L 30 sec 0 0 1 min 0 0 2 min 0 0 3min 0 0 4 min 0 0 5 min 0 0 10 min 0 0

Table 44 summarizes the data obtained in the tests using respectively1,3-dibromo-5,5-dimethylhydantoin (DBDMH) andN,N-bromochloro-5,5-dimethylhydantoin (BCDMH) and in which themicroorganism in each case was Enterococcus faecium. Table 44 shows that1,3-dibromo-5,5-dimethylhydantoin passed the test at one milligram ofbromine, as Br₂, per liter of water, as evidenced by the complete killwithin 30 seconds, whereas N,N′-bromochloro-5,5-dimethylhydantoinrequired two milligrams of bromine, as Br₂, per liter of water toachieve complete kill within 30 seconds. TABLE 44 EFFECTIVENESS AGAINSTENTEROCOCCUS FAECIUM Log₁₀ CFU Log₁₀ CFU Concentration Recovered UsingRecovered Using mg/L as Br₂ Contact Time DBDMH BCDMH 0.5 mg/L 30 sec4.32 >4.48 1 min 2.36 3.53 2 min 0.00 2.63 3 min 0.00 0.00 4 min 0.000.00 5 min 0.00 0.00 10 min 0.00 0.00 1.0 mg/L 30 sec 0.00 >4.48 1 min0.00 2.38 2 min 0.00 0.00 3 min 0.00 0.00 4 min 0.00 0.00 5 min 0.000.00 10 min 0.00 0.00 2.0 mg/L 30 sec 0.00 0.00 1 min 0.00 0.00 2 min0.00 0.00 3 min 0.00 0.00 4 min 0.00 0.00 5 min 0.00 0.00 10 min 0.000.00

Table 45 summarizes test results performed at MBEC Biofilm Technologies,Inc., Calgary, Canada on the effectiveness of various biocides onbiofilm removal. The test procedure, developed at the University ofCalgary, utilizes a device which allows the growth of 96 identicalbiofilms under carefully controlled conditions. The device consists of atwo-part vessel comprised of an upper plate containing 96 pegs thatseals against a bottom plate. The bottom plate can consist of either atrough (for biofilm growth) or a standard 96-well plate (for biocidechallenge). The biofilms develop on the 96 pegs. The device has beenused as a general method for evaluating the efficacy of antibiotics andbiocides towards biofilms. See in this connection H. Ceri, et al., “TheMBEC Test: A New In Vitro Assay Allowing Rapid Screening for AntibioticSensitivity of Biofilm”, Proceedings of the ASM, 1998, 89, 525; Ceri, etal., “Antifungal and Biocide Susceptibility testing of Candida Biofilmsusing the MBEC Device”, Proceedings of the Interscience Conference onAntimicrobial Agents and Chemotherapy, 1998, 38, 495; and H. Ceri, etal., “The Calgary Biofilm Device: A New Technology for the RapidDetermination of Antibiotic Susceptibility of Bacterial Biofilms”,Journal of Clinical Microbiology, 1999, 37, 1771-1776.

Six biocide systems were evaluated using the above test procedure andtest equipment. Five of these systems were oxidizing biocides, viz.,chlorine (from NaOCl), halogen (from NaOCl+NaBr), halogen (from BCDMH),bromine (from DBDMH), and chlorine (from trichloroisocyanuric acid), allexpressed as bromine as Br₂ in mg/L, so that all test results wereplaced on the same basis. The sixth biocide was glutaraldehyde, anon-oxidizing biocide.

These biocide systems were used to challenge biofilms of Pseudomonasaeruginosa (ATCC 15442). This is a Gram (−) bacterium which isubiquitous in microbiological slimes found in many water systems. See inthis connection J. W. Costerton and H. Anwar, “Pseudomonas aeruginosa:The Microbe and Pathogen”, in Pseudomonas aeruginosa Infections andTreatment, A. L. Baltch and R. P. Smith editors, Marcel Dekkerpublishers, New York, 1994. In the field of poultry processing, S.Notermans, J. Dormans, and G. C. Mead, Biofouling, 1991, Vol. 5, pages21-36, report observation of biofilms in poultry slaughter houses by useof scanning electron microscopy.

In Table 45 the MBEC (minimum biofilm eradication concentration) resultspresented are for the one-hour biocide contact time used in the test.The values given for the halogen containing biocides are expressed interms of mg/L of bromine as Br₂. The data on the glutaraldehyde is interms of mg/L as active ingredient. The data indicate that the DBDMH wasmore effective than any of the other biocides tested under theseconditions with an MBEC of 1.4 mg/L of bromine, as Br₂. In fact, onlyslightly more than one-half as much bromine from DBDMH was required toremove the biofilm as compared to the total halogen, expressed as Br₂,that was required from BCDMH. TABLE 45 EFFECTIVENESS AGAINST PSEUDOMONASAERUGINOSA BIOFILM Biocide System MBEC MBEC, avg. Chlorine (from NaOCl)5.0, 2.5 3.8 Halogen (from NaOCl + NaBr) 2.5, 2.5 2.5 Halogen (fromBCDMH) 2.5, 2.5 2.5 Bromine (from DBDMH) 1.4, 1.4 1.4 Chlorine 2.6, 1.32.0 (from Trichloroisocyanuric acid) Glutaraldehyde 50, 50 50

In another group of tests, the results of which are depicted in FIGS. 5through 10, several bromine-based microbiocides of this invention wereutilized in tests illustrating their effectiveness in eradicating orcontrolling Heterotrophic Plate Count bacteria i.e., a mixture ofnaturally-occurring pathogenic bacteria of various unidentified species.These bacteria were challenged both in the form of biofilms and inplanktonic form.

The experimental conditions utilized in these tests involved use of anapparatus consisting of three parallel transparent PVC sampling pipes.These pipes were used for collection of biofilm (i.e., sessile orsurface attached) bacteria samples; one as control pipe, one for arelatively low biocide concentration and the third for a higher biocideconcentration. The biocide challenge in each case was divided into threephases. First was a 14-day inoculation. Next was a 48-hour disinfectionperiod. Finally a 2-week recovery period was provided. The biocide undertest was slug-dosed and during the first hour of exposure, theconcentration was adjusted to achieve the desired concentration level.

The source of the naturally-grown heterotrophic plate count (HPC)bacteria was sediment and associated water collected from therecirculating hot water system of a hospital. Filter cartridges wereinserted into the hospital water system and after about two months asuitable amount of sediment had accumulated on the filters. Thecollected filter/water suspension was then harvested for culturing. Theinoculum for the biocide challenge experiments consisted ofdechlorinated tap water, HPC-cultured stock solution, and a nutrientsupplement solution. The inoculum was incubated at 37° C. for 14-daysprior to the start of the test. The inoculum along with additionaldechlorinated tap water was introduced into the apparatus composed ofthe three parallel transparent PVC sampling pipes. This mixture wasrecirculated throughout the apparatus intermittently at the rate of 3.2gallons per minute for 14-days to produce a consistent biofilm andplanktonic HPC bacteria population.

Samples of these bacteria were collected at the end of the 14-dayinoculation period before the biocide challenge. In each test, the HPCbacteria was then challenged with a specified level of a bromine-basedbiocide, and samples were taken at 1, 2, 3, 12, and 48-hour intervals.These samples were taken by swabbing the inner surface of a premeasuredsection (length, 17/32 inch) of the transparent PVC sampling pipe. Theswabs were vortexed for 1 minute in 5 mL of deionized water with 0.1 mLof a neutralizer (to remove residual bromine) before plating.Concurrently, water samples were taken for enumeration of the planktonicHPC bacteria.

After the 48-hour biocide challenge period, the procedure involvedproviding the 2-week recovery period. The purpose of providing thisrecovery period was to determine how quickly the viable HPC bacteriathat were still present repopulated both the biofilm and, in planktonicform, the recirculating water. Thus, the recirculating water was drainedfrom the test apparatus and the apparatus was refilled withheat-sterilized tap water which was also allowed to recirculateintermittently as before. After 7 and 14 days the apparatus wasresampled and biofilm and planktonic HPC bacteria were enumerated in thesame manner as done previously.

The results of these test are presented in graphical form in the FIGS. 5through 10. In the tests of FIG. 5 the active bromine species derivedfrom sulfamate-stabilized bromine chloride (Stabrom® 909 biocide,Albemarle Corporation) were employed respectively at 0.5 ppm and at 2ppm, both as bromine, to challenge biofilm-associated HPC bacteria. Inaddition a control was carried out in the same manner except that nobiocide was applied. It can be seen that at the higher bromineconcentration, within three hours almost 99% of the biofilm-associatedHPC bacteria were eradicated, whereas at 0.5 ppm as bromine, around 95%of the HPC bacteria were eradicated. It can also be seen that at bothlevels of active bromine concentration, very little recovery of thebiofilm HPC bacteria occurred during the 48-hour biocide challengeperiod. Furthermore, even after the full two-week recovery period, theHPC biofilm bacteria had still not reestablished their originalpopulation level.

In FIG. 6 the active bromine species used in the tests and theirconcentrations were the same as in FIG. 5, and a control was used.However, in these tests the HPC bacteria were in planktonic form. It canbe seen that at the higher bromine concentration, within three hoursover 90% of the planktonic HPC bacteria were eradicated, and at 0.5 ppmas bromine, approx-imately 85% of the planktonic HPC bacteria wereeradicated. These test results also indicate that even at these lowlevels of active bromine, the planktonic HPC bacteria were not able toreestablish populations equal to their original levels during the 2-weekrecovery period.

The results depicted in FIG. 7 involved use of higher concentrations ofthe active bromine species derived from sulfamate-stabilized brominechloride (Stabrom® 909 biocide) than the tests of FIG. 5. In particular,this microbiocide was employed respectively at 4 ppm and at 10 ppm, bothas bromine, to challenge biofilm-associated HPC bacteria. In addition, acontrol was carried out in the same manner except that no biocide wasapplied. It can be seen that at the higher bromine concentration, withinthree hours almost 99.9% of the biofilm-associated HPC bacteria wereeradicated. At 4 ppm as bromine, almost 99% of the HPC bacteria wereeradicated within three hours. It can also be seen that at both levelsof active bromine concentration, very little recovery of the biofilm HPCbacteria occurred during the 48-hour biocide challenge period.Furthermore, even after the full two-week recovery period, the HPCbiofilm bacteria had still not reestablished populations close to theiroriginal levels.

In FIG. 8 the active bromine species used and their concentrations werethe same as in FIG. 7, and a control was used. However, in these teststhe HPC bacteria were in planktonic form. It can be seen that at bothbromine concentration, within three hours over 99% of the planktonic HPCbacteria were eradicated. It can also be seen that within the 48-hourbiocide challenge period, recovery of the very small amounts of theviable planktonic HPC bacteria that still remained had hardly begun tooccur in either of the tests in which the bromine biocide was used.These test results also indicate that for the planktonic HPC bacteria toreestablish populations close to their original levels, a recoveryperiod of substantially more than two weeks would be required.

The test results depicted in FIG. 9 involved use of1,3-dibromo-5,5-dimethylhydantoin (Albrom® 100 biocide, AlbemarleCorporation) as the source of active bromine species. This microbiocidewas used in these tests at levels of 0.5 ppm and 5 ppm as bromine tochallenge biofilm-associated HPC bacteria. Also, a control was carriedout in the same manner except that no biocide was applied. It can beseen from FIG. 9 that at the higher bromine concentration, within twelvehours almost 99.9% of the HPC bacteria were eradicated. At 0.5 ppm asbromine, over 99% of the HPC bacteria were eradicated within threehours. It can also be seen that within the 48-hour biocide challengeperiod, the very small amounts of the viable HPC biofilm that stillremained were beginning to recover in both tests in which the brominebiocide was used. These test results also indicate that for the HPCbacteria to reestablish populations close to their original levels, arecovery period of substantially greater than two weeks would have beenrequired.

In the tests of FIG. 10 the active bromine species used and theirconcentrations were the same as in FIG. 9, and a control was used.However, in these tests the HPC bacteria were in planktonic form. It canbe seen that at the higher bromine concentration, almost 99.99% of theplanktonic HPC bacteria were eradicated within twelve hours. At 0.5 ppmas bromine and within three hours, almost 99% of the planktonic HPCbacteria were eradicated. It can also be seen that within the 48-hourbiocide challenge period, the very small amounts of the viableplanktonic HPC bacteria that still remained were beginning to recover inboth tests in which the bromine biocide was used. These test resultsalso indicate that for the planktonic HPC bacteria to reestablishpopulations close to their original levels, a recovery period of morethan two weeks would have been required.

In the practice of this invention, combinations of different sanitizingsteps using different microbiocidal agents, at least one of which is amicrobiocide of this invention, preferably one or more bromine-basedmicrobiocidal agents of this invention, can prove useful. For example, amicrobiocide of this invention, preferably a bromine-based microbiocideof this invention, can be applied to or contacted with various surfacesassociated with the poultry processing such as conduits, tanks (e.g.,the scalding tank(s), chill tank(s), conveyor belts or conveyor lines,and the poultry carcasses themselves can be treated with anantimicrobial agent such as solutions or gels containing carboxylicacids (e.g., acetic or lactic acid) and/or peroxycarboxylic acids, suchas peracetic acid, peroxyoctanoic acid, peroxydecanoic acid, or thelike. Use of such carboxylic acids is described for example in U.S. Pat.No. 6,113,963. The result of such combined operations is highlyeffective sanitization. In fact, it is contemplated that thiscombination of operations will result in a greater extent ofmicrobiological eradication than has been generally achievableheretofore, especially when the bromine-based biocide used is1,3-dibromo-5,5-dimethylhydantoin and the carboxylic acid used isperacetic acid. Indeed the combined effect of these microbiocides may besynergistic.

Another microbiocide which can be utilized in combined operationspursuant to this invention is trisodium phosphate, a material whichaccording to Capita et al., Meat Science, 2000, 55 (4), 471-474, hasbeen approved by the USDA as an aid to eliminate Salmonella on rawpoultry carcasses. In the combined operations trisodium phosphate isapplied to the poultry carcasses, and one or more of the microbiocidesof this invention, preferably one or more of the bromine-basedmicrobiocides of this invention, are utilized in sanitizing theequipment, instruments, and/or apparatus associated with the processingof the poultry. Also pursuant to this invention the combined operationscan utilize chlorine dioxide treatments along with use of themicrobiocides of this invention. Smith, Meat Processing, 1996, 35(10),47 indicates that chlorine dioxide had been approved by the US FDA foruse in poultry processing water, and in the practice of this inventionone or more microbiocides of this invention, preferably one or more ofthe bromine-based microbiocides of this invention, are utilized insanitation of various items of equipment, instruments, and/or apparatusutilized in the processing of the poultry, and chlorine dioxide is usedto sanitize at least some of the poultry processing water.

Another way by which combined operations pursuant to this invention canbe carried out involves administering to the digestive tract of thepoultry a suitable biological pathogen-control agent, such as byincluding such biological agent in the drinking water for the fowl, oron or in the feed for the fowl. Illustrative biological pathogen-controlagents which may be used in this manner include certain strains of E.coli described in U.S. Pat. No. 6,083,500. Thus in the practice of thisinvention, such a biological pathogen-control agent is provided to thefowl for consumption by drinking and/or eating, and amicrobiocidally-effective amount of an aqueous solution of at least onemicrobiocide of this invention, which preferably is at least onebromine-based microbiocide of this invention, is used in disinfecting orsanitizing equipment, instruments, apparatus, and/or water used in theprocessing of poultry, and/or of carcasses and/or parts of poultryresulting from the processing of poultry.

Still another combined operation involves (i) treating the carcasses ofthe fowl with immobilized lactoferrin antimicrobial agents as describedin U.S. Pat. No. 6,172,040 B1 and (ii) disinfecting or sanitizing all ora portion of the equipment, instruments, apparatus, and/or water used inthe processing of poultry by contacting the same with amicrobiocidally-effective amount of an aqueous solution of at least onemicrobiocide of this invention, which preferably is at least onebromine-based microbiocide of this invention.

Automated dispensing equipment suitable for use in dispensing themicrobiocides of this invention has been described in the literature andto at least some extent is available in the marketplace. For a referenceto such equipment, see for example U.S. Pat. No. 5,683,724 wherein anautomated dispensing system is described.

While chemists understand what is meant by “aqueous” in connection witha solution or medium or the like, it is probably desirable to state forthe benefit of those lawyers who may make it a profession to pettifogover every word someone uses, just what “aqueous” means. The adjective“aqueous” means that the solution or medium or whatever other noun theadjective modifies, can be water whether highly purified or of ordinarypurity such as emanates from the faucet. Since we are dealing withprocessing of food, it stands to reason that one would not use sewerwater or water containing lethal doses of poisons such as cyamide.Besides naturally-occurring trace impurities that may be present in,say, potable water in general, such as ordinary well water or municipalwater, the adjective “aqueous” also permits the presence in the water ofdissolved salts that are formed in the course of forming a bromine-basedmicrobiocide in the water, e.g., by reaction between bromine chlorideand sodium sulfamate in an overbased aqueous solution. In addition,“aqueous” permits the presence of small amounts of innocuousnon-harmful, water-soluble organic solvents such as ethyl alcohol whichcan be used as a solvent for the 1,3-dihalo-5,5-dialkylhydantoin(s).Also “aqueous” permits the presence in the water of the amount of thehalogen-based microbiocide itself to the extent that it may dissolve inthe water, plus any dissolved reactant(s) that may remain after thereaction. Also the water may contain a few atoms that may dissolve fromthe vessel in which the reaction takes place, plus air-borne impuritiesthat may find their way into the water. The point here is that the term“aqueous” does not restrict the medium or solvent to absolutely purewater—the aqueous solution or medium or the like can contain what wouldnormally be present and/or reasonably be expected to be present in itunder the particular circumstances involved when employing ordinarycommon sense.

Compounds referred to by chemical name or formula anywhere in thisdocument, whether referred to in the singular or plural, are identifiedas they exist prior to coming into contact with another substancereferred to by chemical name or chemical type (e.g., another component,a solvent, or etc.). It matters not what chemical changes, if any, takeplace in the resulting mixture or solution, as such changes are thenatural result of bringing the specified substances together under theconditions called for pursuant to this disclosure. As an example, thephase “solution of at least one 1,3-dihalo-5,5-dialkylhydantoin” andphrases of similar import signify that just before being brought intocontact with an aqueous medium such as water, the at least one1,3-dihalo-5,5-dialkylhydantoin referred to was the specified1,3-dihalo-5,5-dialkylhydantoin. The phrase thus is a simple, clear wayof referring to the solution, and it is not intended to suggest or implythat the chemical exists unchanged in the water. The transformationsthat take place are the natural result of bringing these substancestogether, and thus need no further elaboration.

Also, even though the claims may refer to substances in the presenttense (e.g., “comprises”, “is”, etc.), the reference is to the substanceas it exists at the time just before it is first contacted, blended ormixed with one or more other substances in accordance with the presentdisclosure.

Except as may be expressly otherwise indicated, the article “a” or “an”if and as used herein is not intended to limit, and should not beconstrued as limiting, the description or a claim to a single element towhich the article refers. Rather, the article “a” or “an” if and as usedherein is intended to cover one or more such elements, unless the textexpressly indicates otherwise.

All documents referred to herein are incorporated herein by reference intoto as if fully set forth in this document.

This invention is susceptible to considerable variation within thespirit and scope of the appended claims.

1. A method of providing microbiological control in poultry processing,which method comprises disinfecting with a halogen-based microbiocide A)carcasses and/or other parts of poultry resulting from such processing,and B) at least one of (1), (2), (3), (4), where (1), (2), (3), (4) are(1) equipment used in the processing of poultry, (2) instruments used inthe processing of poultry, (3) apparatus used in the processing ofpoultry, (4) water used in the processing of poultry; wherein saidhalogen-based microbiocide is (I) an aqueous microbiocidal solution ofone or more active halogen species, which solution is a derivativeproduct in an aqueous medium of (a) bromine, chlorine, or brominechloride, or any two or all three thereof, and (b) a water-solublesource of sulfamate anion; or (II) an aqueous microbiocidal solution ofone or more active halogen species, which solution is a derivativeproduct in an aqueous medium of at least one1,3-dihalo-5,5-dialkylhydantoin in which one of the halogen atoms is achlorine atom and the other is a chlorine or bromine atom, and in whicheach of the alkyl groups, independently, contains in the range of 1 toabout 4 carbon atoms; or (III) an aqueous microbiocidal solution of oneor more active halogen species, which solution is a derivative productin an aqueous medium of at least one 1,3-dibromo-5,5-dialkylhydantoin inwhich one of the alkyl groups is a methyl group and the other alkylgroup contains in the range of 1 to about 4 carbon atoms; or (IV) anytwo or more of (I), (II), and (III).
 2. The method of claim 1 whereinthe microbiocide used comprises a microbiocidal amount of an aqueousmicrobiocidal solution of one or more active halogen species, whichsolution is a derivative product in an aqueous medium of (a) bromine,chlorine, or bromine chloride, or any two or all three thereof, and (b)a water-soluble source of sulfamate anion.
 3. The method of claim 1wherein the microbiocide used comprises a microbiocidal amount of anaqueous microbiocidal solution of one or more active halogen species,which solution is a derivative product in an aqueous medium of (a)bromine or bromine chloride, or both, and (b) a water-soluble source ofsulfamate anion.
 4. The method of claim 1 wherein the microbiocide usedcomprises an aqueous microbiocidal solution containing amicrobiocidally-effective amount of one or more active halogen species,which solution is a derivative product in an aqueous medium of at leastone 1,3-dihalo-5,5-dialkylhydantoin in which one of the halogen atoms isa chlorine atom and the other is a chlorine or bromine atom, and inwhich each of the alkyl groups, independently, contains in the range of1 to about 4 carbon atoms.
 5. The method of claim 1 wherein themicrobiocide used comprises an aqueous microbiocidal solution containinga microbiocidally-effective amount of one or more active halogenspecies, which solution is a derivative product in an aqueous medium ofat least one N,N′-bromochloro-5,5-dialkylhydantoin in which each of thealkyl groups, independently, contains in the range of 1 to about 4carbon atoms.
 6. The method of claim 1 wherein the microbiocide usedcomprises an aqueous microbiocidal solution containing amicrobiocidally-effective amount of one or more active halogen species,which solution is a derivative product in an aqueous medium ofN,N′-bromochloro-5,5-dimethylhydantoin.
 7. The method of claim 1 whereinthe microbiocide used comprises an aqueous microbiocidal solutioncontaining a microbiocidally-effective amount of one or more activehalogen species, which solution is a derivative product in an aqueousmedium of N,N′-bromochloro-5,5-dimethylhydantoin and1,3-dichloro-5-ethyl-5-methylhydantoin.
 8. The method of claim 1 whereinthe microbiocide used comprises an aqueous microbiocidal solutioncontaining a microbiocidally-effective amount of one or more activehalogen species, which solution is a derivative product in an aqueousmedium of at least one 1,3-dibromo-5,5-dialkylhydantoin in which one ofthe alkyl groups in a methyl group and the other alkyl group contains inthe range of 1 to about 4 carbon atoms.
 9. The method of claim 1 whereinthe microbiocide used comprises an aqueous microbiocidal solutioncontaining a microbiocidally-effective amount of one or more activehalogen species, which solution is a derivative product in an aqueousmedium of 1,3-dibromo-5,5-dimethylhydantoin and1,3-dibromo-5-ethyl-5-methylhydantoin.
 10. The method of claim 1 whereinthe microbiocide used comprises an aqueous microbiocidal solutioncontaining a microbiocidally-effective amount of one or more activehalogen species, which solution is a derivative product in an aqueousmedium of 1,3-dibromo-5,5-dimethylhydantoin.
 11. The method of any ofclaims 1 to 10, both inclusive, wherein A) said carcasses and/or otherparts of poultry resulting from such processing, and B) at least one ofsaid (1), (2), (3), (4), being disinfected have therein or thereon atleast one of Escherichia coli, Salmonella enteritidis, Salmonellatyphimurim, Campylobacter jejuni, Campylobacter coli, Campylobacterlari, Listeria monocytogenes, Pseudomonas fluorescens, Pseudomonasaeruginosa, Enterococcus faecium, and Staphylococcus aureus.